Formulation for oral administration of apoptosis promoter

ABSTRACT

An orally deliverable pharmaceutical composition comprises as a sole or first active ingredient a compound of Formula I defined herein or a pharmaceutically acceptable salt thereof, for example ABT-263 free base or ABT-263 bis-HCl salt, dispersed, in a free base equivalent amount of at least about 2.5% by weight of the composition, in a pharmaceutically acceptable carrier; wherein said active ingredient is in solid-state form and/or the composition further comprises, dispersed in the carrier, a pharmaceutically acceptable heavier-chalcogen antioxidant in an amount effective to inhibit oxidation of the active ingredient at a thioether linkage thereof. The composition is suitable for oral administration to a subject in need thereof for treatment of a disease characterized by overexpression of one or more anti-apoptotic Bcl-2 family proteins, for example cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority benefit of U.S. Provisional ApplicationSer. No. 61/174,299 filed on Apr. 30, 2009, Ser. No. 61/174,318 filed onApr. 30, 2009, Ser. No. 61/185,105 filed on Jun. 8, 2009, Ser. No.61/185,130 filed on Jun. 8, 2009, Ser. No. 61/218,281 filed on Jun. 18,2009, Ser. No. 61/289,254 filed on Dec. 22, 2009, and Ser. No.61/289,289 filed on Dec. 22, 2009.

Cross-reference is made to the following co-filed U.S. applicationscontaining subject matter related to the present application: Ser. No.12/______ titled “Lipid formulation of apoptosis promoter”, which claimspriority benefit of U.S. provisional application Ser. No. 61/174,245filed on Apr. 30, 2009; Ser. No. 12/______ titled “Salt of ABT-263 andsolid-state forms thereof”, which claims priority benefit of U.S.provisional application Ser. No. 61/174,274 filed on Apr. 30, 2009; Ser.No. 12/______ titled “Stabilized lipid formulation of apoptosispromoter”, which claims priority benefit of above-referenced U.S.provisional application Ser. No. 61/174,299 and Ser. No. 61/289,254; andSer. No. 12/______ titled “Solid oral formulation of ABT-263”, whichclaims priority benefit of above-referenced U.S. provisional applicationSer. No. 61/174,318.

The entire disclosure of each of the above applications is incorporatedherein by reference.

FIELD OF THE INVENTION

The present invention relates to pharmaceutical compositions comprisingan apoptosis-promoting agent, for example ABT-263, and to methods of usethereof for treating diseases characterized by overexpression ofanti-apoptotic Bcl-2 family proteins. More particularly the inventionrelates to such compositions that exhibit improved stability andadequate oral bioavailability, and to oral dosage regimens foradministration of such a composition to a subject in need thereof.

BACKGROUND OF THE INVENTION

Evasion of apoptosis is a hallmark of cancer (Hanahan & Weinberg (2000)Cell 100:57-70). Cancer cells must overcome a continual bombardment bycellular stresses such as DNA damage, oncogene activation, aberrant cellcycle progression and harsh microenvironments that would cause normalcells to undergo apoptosis. One of the primary means by which cancercells evade apoptosis is by up-regulation of anti-apoptotic proteins ofthe Bcl-2 family.

Compounds that occupy the BH3 binding groove of Bcl-2 proteins have beendescribed, for example by Bruncko et al. (2007) J. Med. Chem.50:641-662. These compounds have includedN-(4-(4-((4′-chloro-(1,1′-biphenyl)-2-yl)methyl)piperazin-1-yl)benzoyl)-4-(((1R)-3-(dimethylamino)-1-((phenylsulfanyl)methyl)propyl)amino)-3-nitrobenzenesulfonamide,otherwise known as ABT-737, which has the formula:

ABT-737 binds with high affinity (<1 nM) to proteins of the Bcl-2 family(specifically Bcl-2, Bcl-X_(L) and Bcl-w). It exhibits single-agentactivity against small-cell lung cancer (SCLC) and lymphoidmalignancies, and potentiates pro-apoptotic effects of otherchemotherapeutic agents. ABT-737 and related compounds, and methods tomake such compounds, are disclosed in U.S. Patent ApplicationPublication No. 2007/0072860 of Bruncko et al.

More recently, a further series of compounds has been identified havinghigh binding affinity to Bcl-2 family proteins. These compounds, andmethods to make them, are disclosed in U.S. Patent ApplicationPublication No. 2007/0027135 of Bruncko et al. (herein “the '135publication”), incorporated by reference herein in its entirety, and canbe seen from their formula to be structurally related to ABT-737.

One compound, identified as “Example 1” in the '135 publication, isN-(4-(4-((2-(4-chlorophenyl)-5,5-dimethyl-1-cyclohex-1-en-1-yl)methyl)piperazin-1-yl)benzoyl)-4-(((1R)-3-(morpholin-4-yl)-1-((phenylsulfanyl)methyl)propyl)amino)-3-((trffluoromethyl)sulfonyl)benzene-sulfonamide,otherwise known as ABT-263. This compound has a molecular weight of974.6 g/mol and has the formula:

The '135 publication states that while inhibitors of Bcl-2 familyproteins previously known may have either potent cellular efficacy orhigh systemic exposure after oral administration, they do not possessboth properties. A typical measure of cellular efficacy of a compound isthe concentration eliciting 50% cellular effect (EC₅₀). A typicalmeasure of systemic exposure after oral administration of a compound isthe area under the curve (AUC) resulting from graphing plasmaconcentration of the compound versus time from oral administration.Previously known compounds, it is stated in the '135 publication, have alow AUC/EC₅₀ ratio, meaning that they are not orally efficacious. Bycontrast, compounds provided therein are stated to demonstrate enhancedproperties with respect to cellular efficacy and systemic exposure afteroral administration, resulting in a AUC/EC₅₀ ratio significantly higherthan that of previously known compounds.

ABT-263 binds with high affinity (<1 nM) to Bcl-2 and Bcl-X_(L) and isbelieved to have similarly high affinity for Bcl-w. Its AUC/EC₅₀ ratiois reported in the '135 publication as 56, more than an order ofmagnitude greater than that reported for ABT-737 (4.5). Fordetermination of AUC according to the '135 publication, each compoundwas administered to rats in a single 5 mg/kg dose by oral gavage as a 2mg/ml solution in a vehicle of 10% DMSO (dimethyl sulfoxide) in PEG-400(polyethylene glycol of average molecular weight about 400).

Oral bioavailability (as expressed, for example, by AUC after oraladministration as a percentage of AUC after intravenous administration)is not reported in the '135 publication, but can be concluded therefromto be, at least in a rat model, substantially greater for ABT-263 thanfor ABT-737, when administered in PEG-400/DMSO solution.

Various solutions to the challenge of low oral bioavailability have beenproposed in the art. For example, U.S. Pat. No. 5,645,856 to Lacy et al.proposes formulating a hydrophobic drug with (a) an oil, (b) ahydrophilic surfactant and (c) a lipophilic surfactant thatsubstantially reduces an inhibitory effect of the hydrophilic surfactanton in vivo lipolysis of the oil, such lipolysis being said to be afactor promoting bioavailability of the drug. Among numerous classes ofhydrophilic surfactants listed are phospholipids such as lecithins.

U.S. Pat. No. 6,267,985 to Chen & Patel is directed, inter alia, to apharmaceutical composition comprising (a) a triglyceride, (b) a carriercomprising at least two surfactants, one of which is hydrophilic, and(c) a therapeutic agent capable of being solubilized in thetriglyceride, the carrier or both. It is specified therein that thetriglyceride and the surfactants must be present in amounts providing aclear aqueous dispersion when the composition is mixed with an aqueoussolution under defined conditions. Among extensive separate lists ofexemplary ingredients, mention is made of “glyceryltricaprylate/caprate” as a triglyceride, and phospholipids includingphosphatidylcholine as surfactants.

U.S. Pat. No. 6,451,339 to Patel & Chen mentions disadvantages ofpresence of triglycerides in such compositions, and proposes otherwisesimilar compositions that are substantially free of triglycerides, butthat likewise provide clear aqueous dispersions.

U.S. Pat. No. 6,309,663 to Patel & Chen proposes pharmaceuticalcompositions comprising a combination of surfactants said to enhancebioabsorption of a hydrophilic therapeutic agent. Phospholipids such asphosphatidylcholine are again listed among exemplary surfactants.

U.S. Pat. No. 6,464,987 to Fanara et al. proposes a fluid pharmaceuticalcomposition comprising an active substance, 3% to 55% by weight ofphospholipid, 16% to 72% by weight of solvent, and 4% to 52% by weightof fatty acid. Compositions comprising Phosal 50 PG™ (primarilycomprising phosphatidylcholine and propylene glycol), in some casestogether with Phosal 53 MCT™ (primarily comprising phosphatidylcholineand medium chain triglycerides), are specifically exemplified. Suchcompositions are said to have the property of gelling instantaneously inpresence of an aqueous phase and to allow controlled release of theactive substance.

U.S. Pat. No. 5,538,737 to Leonard et al. proposes a capsule containinga water-in-oil emulsion wherein a water-soluble drug salt is dissolvedin the water phase of the emulsion and wherein the oil phase comprisesan oil and an emulsifying agent. Among oils mentioned are medium chaintriglycerides; among emulsifying agents mentioned are phospholipids suchas phosphatidylcholine. Phosal 53 MCT™, which containsphosphatidylcholine and medium chain triglycerides, is reportedly usedaccording to various examples therein.

U.S. Pat. No. 5,536,729 to Waranis & Leonard proposes an oralformulation comprising rapamycin, at a concentration of about 0.1 toabout 50 mg/ml, in a carrier comprising a phospholipid solution. It isstated therein that a preferred formulation can be made using Phosal 50PG™ as the phospholipid solution. An alternative phospholipid solutionmentioned is Phosal 50 MCT™.

U.S. Pat. No. 5,559,121 to Harrison et al. proposes an oral formulationcomprising rapamycin, at a concentration of about 0.1 to about 100mg/ml, in a carrier comprising N,N-dimethylacetamide and a phospholipidsolution. Examples of the more preferred embodiments are shown to beprepared using Phosal 50 PG™. An alternative phospholipid solutionmentioned is Phosal 50 MCT™.

U.S. Patent Application Publication No. 2007/0104780 of Lipari et al.discloses that a small-molecule drug (defined therein as havingmolecular weight, excluding counterions in the case of salts, notgreater than about 750 g/mol, typically not greater than about 500g/mol) having low water solubility can be formulated as a solution in asubstantially non-aqueous carrier comprising at least one phospholipidand a pharmaceutically acceptable solubilizing agent. The solution, whenmixed with an aqueous phase, is said to form a non-gelling,substantially non-transparent liquid dispersion. Illustratively,formulations ofN-(4-(3-amino-1H-indazol-4-yl)phenyl)-N′-(2-fluoro-5-methylphenyl)urea(the protein tyrosine kinase inhibitor ABT-869) comprising Phosal 53MCT™ and other ingredients are described therein.

Recently, Tse et al. (2008) Cancer Res. 68(9):3421-3428, reported insupplementary data thereto that, in a dog model, oral bioavailability ofan ABT-263 solution in PEG-400/DMSO was 22.4%, and that of an ABT-263solution in 60% Phosal™ PG (phosphatidylcholine+propylene glycol), 30%PEG-400 and 10% ethanol was 47.6%.

At the time of the present invention, however, the art was silent as towhether compounds of the '135 publication such as ABT-263 havesufficient chemical stability to permit formulation in pharmaceuticalcompositions suitable as storable, transportable materials of commerceas opposed to extemporaneously prepared solutions. Further, the art gaveno indication as to whether, if such compositions could be made, theywould have acceptable oral bioavailability. Still further, the art wassilent as to whether, if such compositions could be made havingacceptable oral bioavailability, they could have a concentration ofactive ingredient sufficient to provide therapeutically effective dailydosing without the need to swallow an unacceptably large volume ofliquid or an unacceptably large number of discrete solid dosage formssuch as capsules or tablets.

Oxidation reactions represent an important degradation pathway ofpharmaceuticals, especially when formulated in solution. A large body ofinformation is available on oxidative mechanisms, but relatively fewstudies have been performed with specific drugs. Hovorka & Schoneich(2001) J. Pharm. Sci. 90:253-269 have stated that this lack ofpharmaceutically relevant data leads to poor predictive ability withrespect to drug oxidation between manufacture and administration offormulations of oxidizable drugs, and a consequently uninformed, largelyempirical utilization of antioxidants in formulations.

Oxidation can occur by a number of pathways, including uncatalyzedautoxidation of a substrate by molecular oxygen, photolytic initiation,hemolytic thermal cleavage, and metal catalysis. Various functionalgroups show particular sensitivity towards oxidation. In particular,thioethers can degrade via hydrogen abstraction at the α-position to thesulfur atom or by addition of an α-peroxyl radical directly or via aone-electron transfer process, which transforms a sulfide to a sulfine,sulfone, or sulfoxide (Hovorka & Schoneich, supra).

A particular type of disease for which improved therapies are needed isnon-Hodgkin's lymphoma (NHL). NHL is the sixth most prevalent type ofnew cancer in the U.S. and occurs primarily in patients 60-70 years ofage. NHL is not a single disease but a family of related diseases, whichare classified on the basis of several characteristics includingclinical attributes and histology.

One method of classification places different histological subtypes intotwo major categories based on natural history of the disease, i.e.,whether the disease is indolent or aggressive. In general, indolentsubtypes grow slowly and are generally incurable, whereas aggressivesubtypes grow rapidly and are potentially curable. Follicular lymphomasare the most common indolent subtype, and diffuse large-cell lymphomasconstitute the most common aggressive subtype. The oncoprotein Bcl-2 wasoriginally described in non-Hodgkin's B-cell lymphoma.

Treatment of follicular lymphoma typically consists ofbiologically-based or combination chemotherapy. Combination therapy withrituximab, cyclophosphamide, doxorubicin, vincristine and prednisone(R-CHOP) is routinely used, as is combination therapy with rituximab,cyclophosphamide, vincristine and prednisone (RCVP). Single-agenttherapy with rituximab (targeting CD20, a phosphoprotein uniformlyexpressed on the surface of B-cells) or fludarabine is also used.Addition of rituximab to chemotherapy regimens can provide improvedresponse rate and increased progression-free survival.

Radioimmunotherapy agents, high-dose chemotherapy and stem celltransplants can be used to treat refractory or relapsed non-Hodgkin'slymphoma. Currently, there is not an approved treatment regimen thatproduces a cure, and current guidelines recommend that patients betreated in the context of a clinical trial, even in a first-linesetting.

First-line treatment of patients with aggressive large B-cell lymphomatypically consists of rituximab, cyclophosphamide, doxorubicin,vincristine and prednisone (R-CHOP), or dose-adjusted etoposide,prednisone, vincristine, cyclophosphamide, doxorubicin and rituximab(DA-EPOCH-R).

Most lymphomas respond initially to any one of these therapies, buttumors typically recur and eventually become refractory. As the numberof regimens patients receive increases, the more chemotherapy-resistantthe disease becomes. Average response to first-line therapy isapproximately 75%, 60% to second-line, 50% to third-line, and about35-40% to fourth-line therapy. Response rates approaching 20% with asingle agent in a multiple relapsed setting are considered positive andwarrant further study.

Current chemotherapeutic agents elicit their antitumor response byinducing apoptosis through a variety of mechanisms. However, many tumorsultimately become resistant to these agents. Bcl-2 and Bcl-X_(L) havebeen shown to confer chemotherapy resistance in short-term survivalassays in vitro and, more recently, in vivo. This suggests that ifimproved therapies aimed at suppressing the function of Bcl-2 andBcl-X_(L) can be developed, such chemotherapy-resistance could besuccessfully overcome.

Apoptosis-promoting drugs that target Bcl-2 family proteins such asBcl-2 and Bcl-X_(L) are best administered according to a regimen thatprovides continual, for example daily, replenishment of the plasmaconcentration, to maintain the concentration in a therapeuticallyeffective range. This can be achieved by daily parenteral, e.g.,intravenous (i.v.) or intraperitoneal (i.p.) administration. However,daily parenteral administration is often not practical in a clinicalsetting, particularly for outpatients. To enhance utility of anapoptosis-promoting agent, whether in a clinical or community setting,for example as a chemotherapeutic in cancer patients, an orallybioavailable dosage form having sufficient storage-stability not to belimited to extemporaneous preparation would be highly desirable. Such adosage form, and a regimen for oral administration thereof, wouldrepresent an important advance in treatment of many types of cancer,including non-Hodgkin's lymphoma, and would more readily enablecombination therapies with other chemotherapeutics.

SUMMARY OF THE INVENTION

As reported in the '135 publication, oral bioavailability of a dilute (2mg/ml) solution of ABT-263 free base in PEG-400/DMSO in a rat model isaround 20%. Tse et al. (2008), supra, report that a similar solution hascomparable bioavailability of around 20% in other species, including dogand monkey, but that improved bioavailability is obtainable, at least ina dog model, by use of a lipid carrier, namelyphosphatidylcholine/propylene glycol/PEG-400/ethanol. The concentrationsof ABT-263 free base in the PEG-400/DMSO and lipid carriers as tested indogs are not reported by Tse et al., but are disclosed herein to havebeen 5 and 10 mg/ml (approximately 0.5% and 1% by weight) respectively.

Recent U.S. Patent Application Publication No. 2009/0149461 of Krivoshik(“the '461 publication”), incorporated herein by reference in itsentirety without admission that it constitutes prior art to the presentapplication, reports a Phase 1 clinical trial of ABT-263, formulatedextemporaneously as a 25 mg/ml solution in Phosal 53 MCT™ (a proprietaryproduct described hereinafter) and ethanol. It is predicted therein,based on preclinical evidence, that therapeutically effective doses ofABT-263 in human patients will be 200-350 mg/day (see the '461publication at paragraph [0017] bridging pp. 1-2 and paragraph [0032] onp. 3).

Given the variation in individual patients' body weight, therapeuticresponse and tolerance of side-effects, as well as variation inbioavailability of different formulations, a suitable daily dose formost patients is likely to be found in a range of about 50 to about 500mg, more typically about 200 to about 400 mg. Illustratively, to deliverper os 200-400 mg of ABT-263 in the form of a 10 mg/ml (approximately 1%by weight) solution in a lipid carrier requires administration of 20-40ml of solution per day. If encapsulated in easy-to-swallow liquid-filledcapsules, each containing 0.5 ml, this amounts to 40 capsules per day ata 200 mg dose and 80 capsules per day at a 400 mg dose. This is highlyinconvenient for the patient and caregiver, and is likely to result inpoor patient compliance. A 25 mg/ml (approximately 2.5% by weight)ABT-263 concentration, as used in the study reported in the '461publication, represents a minimum threshold for clinical acceptability,requiring daily administration of 8-16 ml of solution, or 16-32 capsuleseach containing 0.5 ml. Further increasing the concentration of activeingredient to provide a less voluminous dosage form, without excessivelysacrificing oral bioavailability, is therefore an important desideratum.However, the physical properties of ABT-263, including its lowsolubility in aqueous and many non-aqueous solvents, make this asignificant technical challenge.

Compounding the difficulty of formulating compounds of the '135publication such as ABT-263, other than as an extemporaneously preparedsolution, is the finding that such compounds are susceptible tooxidation, for example in presence of oxygen or reactive oxygen speciessuch as superoxide, hydrogen peroxide or hydroxyl radicals. The term“extemporaneously prepared” herein means preparation not more than onemonth before, for example not more than one week before, not more thanone day before, or immediately before, administration to a patient inneed thereof. If a formulation is to have acceptable storage-stabilityfor longer than about one month, a solution to the challenge ofoxidative degradation of the active ingredient is required.

The (phenylsulfanyl)methyl group of compounds of the '135 publicationhave a thioether linkage, which is now known to be susceptible tooxidation, for example in presence of oxygen or reactive oxygen speciessuch as superoxide, hydrogen peroxide or hydroxyl radicals. Theabove-referenced '135 publication includes antioxidants in an extensivelist of excipients said to be useful for administering such compounds.

A number of novel and unexpected findings have led, at least in part, tothe present invention. These include the following:

-   -   Lipid solution compositions of compounds of the '135 publication        such as ABT-263 or a salt thereof are, as indicated above,        susceptible to oxidative degradation of the active ingredient.        Not all antioxidants are effective to inhibit this oxidative        degradation. However, it has been found that a particular class        of antioxidants, described herein as “heavier-chalcogen        antioxidants” or “HCAs”, are useful if included in an        antioxidant-effective amount.    -   The requirement to maintain in a physically stable liquid        formulation not only the active ingredient but, additionally, an        HCA in an antioxidant-effective amount can further limit the        choice of liquid carrier, particularly for higher active        ingredient loadings, for example about 50 mg/ml or higher.    -   Compounds of the '135 publication such as ABT-263 or a salt        thereof in solid-state form are typically less susceptible to        oxidative degradation than in solution form. Providing the        carrier also in solid-state form, for example as a polymeric        matrix wherein solid-state active ingredient is dispersed, or as        a dry-blend or granulated mixture of excipients including at        least a diluent and a disintegrant, is therefore another        approach to inhibiting oxidative degradation.    -   Solid dispersion formulations comprising a compound of the '135        publication such as ABT-263 or a salt thereof in an amorphous        form, dispersed in a polymeric matrix, can be prepared at active        ingredient loadings of up to about 25% by weight or even higher.        Such formulations exhibit acceptable resistance to oxidative        degradation and, if they contain a suitable surfactant to        solubilize the active ingredient in gastrointestinal fluid upon        release from the matrix, are found to have acceptable oral        bioavailability in a dog model.    -   Remarkably for such a poorly water-soluble drug, ABT-263 or a        salt thereof formulated as a conventional dry-blend or        granulated mixture with excipients including at least a diluent        and a disintegrant at an active ingredient loading of up to        about 40% by weight or even higher, exhibits generally        acceptable oral bioavailability. Even more remarkably, particle        size reduction is not essential to achieving acceptable bio        availability, although it can provide more rapid release of the        active ingredient.    -   As an alternative liquid formulation, a suspension of        crystalline active ingredient (for this purpose a crystalline        salt such as ABT-263 bis-HCl is preferred) can be prepared in an        aqueous carrier, at ABT-263 free base equivalent concentrations        of at least about 25 mg/ml, for example about 50 mg/ml or        higher, by appropriate selection of surfactant as a suspending        agent. Particle size reduction to provide a D₉₀ not greater than        about 2 μm, for example not greater than about 1 μm, provides a        nanosuspension having remarkably high oral bioavailability,        comparable to that of a lipid solution formulation.

In accordance with these findings, there is now provided an orallydeliverable pharmaceutical composition comprising as a sole or firstactive ingredient a compound of Formula I

where X³ is chloro or fluoro; and

-   -   (1) X⁴ is azepan-1-yl, morpholin-4-yl, 1,4-oxazepan-4-yl,        pyrrolidin-1-yl, —N(CH₃)₂, —N(CH₃)(CH(CH₃)₂),        7-azabicyclo[2.2.1]heptan-7-yl or        2-oxa-5-azabicyclo[2.2.1]hept-5-yl; and R⁰ is

-   -   -   where X⁵ is —CH₂—, —C(CH₃)₂— or —CH₂CH₂—; X⁶ and X⁷ are both            —H or both methyl; and X⁸ is fluoro, chloro, bromo or iodo;            or

    -   (2) X⁴ is azepan-1-yl, morpholin-4-yl, pyrrolidin-1-yl,        —N(CH₃)(CH(CH₃)₂) or 7-azabicyclo[2.2.1]heptan-7-yl; and R⁰ is

-   -   -   where X⁶, X⁷ and X⁸ are as above; or

    -   (3) X⁴ is morpholin-4-yl or —N(CH₃)₂; and R⁰ is

-   -   -   where X⁸ is as above;            or a pharmaceutically acceptable salt thereof, dispersed, in            a free base equivalent amount of at least about 2.5% by            weight of the composition, in a pharmaceutically acceptable            carrier; wherein said active ingredient is in solid-state            form and/or the composition further comprises, dispersed in            the carrier, a pharmaceutically acceptable HCA in an amount            effective to inhibit oxidation of the active ingredient at a            thioether linkage thereof.

In some embodiments, the sole or first active ingredient is ABT-263 or apharmaceutically acceptable salt thereof, for example ABT-263 free baseor ABT-263 bis-hydrochloride salt (ABT-263 bis-HCl).

According to such embodiments, it is preferred that the carrier shouldcomprise excipients selected to provide sufficient bioavailability ofABT-263 to be therapeutically effective for promotion of apoptosis whenorally administered to a non-fasting human subject in need thereof in adaily dosage amount of about 200 to about 400 mg ABT-263 free baseequivalent. “Sufficient bioavailability” in this context can beevidenced, for example, by

-   -   bioavailability of at least about 15% in a non-fasting dog        model;    -   one or both of (a) an ABT-263 AUC₀₋₂₄ of at least about 20        μg·h/ml, and/or (b) an ABT-263 C_(max) of at least about 2.5        μg/ml, in a single-dose non-fasting human pharmacokinetic study        at an ABT-263 free base equivalent dose of about 200 to about        400 mg;    -   a steady-state ABT-263 C_(min) of about 1 to about 5 μg/ml and a        steady-state ABT-263 C_(max) of about 3 to about 8 μg/ml in a        non-fasting human pharmacokinetic study at a daily ABT-263 free        base equivalent dose of about 200 to about 400 mg; or    -   at least substantial bioequivalence in a human pharmacokinetic        study to a prototype extemporaneously prepared formulation that        consists of a 25 mg/ml solution of ABT-263 bis-HCl in a mixture        of 90% phosphatidylcholine+medium chain triglycerides 53/29 and        10% ethanol.

In some embodiments, the carrier is liquid, having the active ingredientand a pharmaceutically acceptable HCA in an antioxidant-effective amountin solution or suspension therein.

In other embodiments, the carrier is solid, having the active ingredientdispersed therein in solid-state form. In such embodiments, presence ofa pharmaceutically acceptable HCA is optional. The term “solid-state”,as used herein to describe a physical form of the active ingredient,includes crystalline, semi-crystalline, amorphous, and solid or glassysolution forms. Crystalline, semi-crystalline and amorphous forms can beessentially solvent-(including water-) free or can take the form ofsolvates or hydrates of the active ingredient.

There is further provided a method for treating a disease characterizedby apoptotic dysfunction and/or overexpression of an anti-apoptoticBcl-2 family protein, comprising orally administering to a subjecthaving the disease a therapeutically effective amount of a compositionas described above. Examples of such a disease include many neoplasticdiseases including cancers. A specific illustrative type of cancer thatcan be treated according to the present method is non-Hodgkin'slymphoma. Another specific illustrative type of cancer that can betreated according to the present method is chronic lymphocytic leukemia.Yet another specific illustrative type of cancer that can be treatedaccording to the present method is acute lymphocytic leukemia, forexample in a pediatric patient.

Additional embodiments of the invention, including more particularaspects of those provided above, will be found in, or will be evidentfrom, the detailed description that follows.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic phase diagram of ABT-263 free base solutions internary “IPT” lipid systems as described in Example 8. The shadedportion of the diagram represents an area of optimized formulationcomposition.

FIG. 2 is a schematic phase diagram of ABT-263 free base solutions internary “IST” lipid systems as described in Example 8. The shadedportion of the diagram represents an area of optimized formulationcomposition.

FIG. 3 is a graphical representation of ABT-263 plasma concentrationover a 24-hour period following oral administration to dogs (non-fastedexcept where otherwise indicated) of a composition of the invention(Formulation 8) and a comparative solution of ABT-263 bis-HCl in a lipidmedium (Formulation C), as described in Example 15.

FIG. 4 is a graphical representation of effects of various surfactantson dissolution rates of solid dispersions containing ABT-263 bis-HCl asdescribed in Example 18.

FIG. 5 is a graphical representation of effects of various surfactantson dissolution rates of solid dispersions containing ABT-263 free baseas described in Example 18.

FIG. 6 is a graphical representation of effects of various polymericcarriers on dissolution rates of solid dispersions containing ABT-263bis-HCl as described in Example 19.

FIG. 7 shows plasma concentration of ABT-263 at different time pointsfollowing oral administration to fasted or fed dogs of an ABT-263bis-HCl solid dispersion formulation containing Span™ 20 as solubilizer,at doses of 50, 100 or 200 mg, as described in Example 23.

FIG. 8 shows plasma concentration of ABT-263 at different time pointsfollowing oral administration to fasted or fed dogs of an ABT-263bis-HCl solid dispersion formulation containing TPGS as solubilizer, atdoses of 50, 100 or 200 mg, as described in Example 23.

FIG. 9 shows plasma concentration of ABT-263 at different time pointsfollowing oral administration to fed dogs of ABT-263 free base orABT-263 bis-HCl solid dispersion formulations containing TPGS only, orTPGS+propylene glycol as plasticizer, at a dose of 50 mg, as describedin Example 24.

FIGS. 10 and 11 show results of an accelerated stability study usingopen dishes, wherein the sulfoxide content of different ABT-263 soliddispersion formulations was determined at different time points, asdescribed in Example 25.

FIGS. 12 and 13 show results of an accelerated stability study usingclosed bottles, wherein the sulfoxide content of different ABT-263 soliddispersion formulations was determined at different time points, asdescribed in Example 25.

FIG. 14 shows release of ABT-263 from tablets containing differentABT-263 solid dispersion formulations, as described in Example 28.

DETAILED DESCRIPTION

The invention is described herein with specific reference to thefollowing embodiments.

In a first composition embodiment, there is provided an orallydeliverable pharmaceutical composition comprising (a) a compound ofFormula I as defined hereinabove, or a pharmaceutically acceptable saltthereof, in a free base equivalent amount of at least about 2.5% byweight of the composition; (b) a pharmaceutically acceptableheavier-chalcogen antioxidant (HCA); and (c) a substantially non-aqueouspharmaceutically acceptable carrier that comprises one or more lipids;wherein said compound and the antioxidant are in solution in thecarrier.

In a second composition embodiment, there is provided an orallydeliverable pharmaceutical capsule comprising a capsule shell havingencapsulated therewithin, in an amount not greater than about 1000 mgper capsule, a liquid solution of a compound of Formula I as definedhereinabove, or a pharmaceutically acceptable salt thereof, in a freebase equivalent amount of at least about 2.5% by weight of the solution,in a substantially non-ethanolic carrier that comprises aspharmaceutically acceptable excipients:

-   -   (a) at least one phospholipid,    -   (b) at least one solubilizing agent for the at least one        phospholipid, selected from the group consisting of glycols,        glycolides, glycerides and mixtures thereof,    -   (c) at least one non-phospholipid surfactant, and    -   (d) a pharmaceutically acceptable HCA.

In a third composition embodiment, there is provided an orallydeliverable liquid pharmaceutical composition comprising an aqueousmedium having suspended therein a solid particulate compound having aD₉₀ particle size not greater than about 3 μm; wherein the compound isof Formula I as defined hereinabove, or a pharmaceutically acceptablesalt thereof, and is present in a free base equivalent amount of atleast about 2.5% by weight of the composition; and wherein the aqueousmedium further comprises at least one pharmaceutically acceptablesurfactant and at least one pharmaceutically acceptable basifying agentin amounts that are effective together to inhibit particle sizeincrease.

In a fourth composition embodiment, there is provided an orallydeliverable solid dispersion comprising, in essentially non-crystalline,for example amorphous, form, a compound of Formula I as definedhereinabove, or a pharmaceutically acceptable salt thereof, in a freebase equivalent amount of at least about 2.5% by weight of thecomposition, dispersed in a solid matrix that comprises (a) apharmaceutically acceptable water-soluble polymeric carrier and (b) apharmaceutically acceptable surfactant.

In a fifth composition embodiment, there is provided an orallydeliverable pharmaceutical dosage form comprising a solid dispersion orsolid solution that comprises (a) a compound of Formula I as definedhereinabove, or a pharmaceutically acceptable salt thereof, in a freebase equivalent amount of at least about 2.5% by weight of thecomposition, (b) at least one pharmaceutically acceptable polymer and(c) at least one pharmaceutically acceptable solubilizer.

In a sixth composition embodiment, there is provided an orallydeliverable pharmaceutical composition comprising (a) a compound ofFormula I as defined hereinabove, or a pharmaceutically acceptable saltthereof, in solid particulate form and in a free base equivalent amountof at least about 2.5% by weight of the composition, and (b) a pluralityof pharmaceutically acceptable excipients including at least a soliddiluent and a solid disintegrant.

Variants of these six composition embodiments will be readily envisionedby one of skill in the art reading the present disclosure, such variantsbeing embraced by the present invention. As indicated above, acomposition of the present invention is, broadly, an orally deliverablepharmaceutical composition comprising as a sole or first activeingredient a compound of Formula I or a pharmaceutically acceptable saltthereof, dispersed, in a free base equivalent amount of at least about2.5% by weight of the composition, in a pharmaceutically acceptablecarrier; wherein said active ingredient is in solid-state form and/orthe composition further comprises, dispersed in the carrier, apharmaceutically acceptable HCA in an amount effective to inhibitoxidation of the active ingredient at a thioether linkage thereof.

Compositions of any of the above embodiments can be used in a method ofthe invention for treating a disease characterized by apoptoticdysfunction and/or overexpression of an anti-apoptotic Bcl-2 familyprotein, for example a neoplastic disease such as cancer. Such a methodcomprises orally administering to a subject having the disease atherapeutically effective amount of a composition as described herein.

A composition of the invention is “orally deliverable”, i.e., adaptedfor oral administration; however, such a composition can be useful fordelivery of the drug to a subject in need thereof by other routes ofadministration, including without limitation parenteral, sublingual,buccal, intranasal, pulmonary, topical, transdermal, intradermal,ocular, otic, rectal, vaginal, intragastric, intracranial, intrasynovialand intra-articular routes.

The terms “oral administration” and “orally administered” herein referto administration to a subject per os (p.o.), that is, administrationwherein the composition is immediately swallowed, for example with theaid of a suitable volume of water or other potable liquid. “Oraladministration” is distinguished herein from intraoral administration,e.g., sublingual or buccal administration or topical administration tointraoral tissues such as periodontal tissues, that does not involveimmediate swallowing of the composition.

A compound of Formula I or salt thereof can be the sole activeingredient in the composition, in which case the compound or salt can beadministered in monotherapy or in combination therapy with one or moreother drugs formulated separately from the compound of Formula I or saltthereof. Alternatively, a compound of Formula I or salt thereof can beaccompanied in the composition by one or more additional drugs, for usein combination therapy. In that case, the compound of Formula I or saltthereof is considered the “first active ingredient” for the purpose ofthe present disclosure.

Therapeutically active compounds, including salts, useful hereintypically have low solubility in water, for example less than about 100μg/ml, in most cases less than about 30 μg/ml. The present invention canbe especially advantageous for drugs that are essentially insoluble inwater, i.e., having a solubility of less than about 10 μg/ml. Examplesof such drugs are include Biopharmaceutics Classification System (BCS)Class IV drug substances that are characterized by low solubility andlow permeability (see “Waiver of in vivo bioavailability andbioequivalence studies for immediate-release solid oral dosage formsbased on a biopharmaceutics classification system”, U.S. Department ofHealth and Human Services, Food and Drug Administration, Center for DrugEvaluation and Research (CDER), August 2000). It will be recognized thataqueous solubility of many compounds is pH-dependent; in the case ofsuch compounds the solubility of interest herein is at a physiologicallyrelevant pH, for example a pH of about 1 to about 8. Thus, in variousembodiments, the drug has a solubility in water, at least at one pointin a pH range from about 1 to about 8, of less than about 100 μg/ml, forexample less than about 30 μg/ml, or less than about 10 μg/ml.Illustratively, ABT-263 has a solubility in water at pH 2 of less than 4μg/ml.

In one embodiment, the composition comprises a compound of Formula I asdefined above, or a pharmaceutically acceptable salt of such a compound.

In a further embodiment, the compound has Formula I where X³ is fluoro.

In a still further embodiment, the compound has Formula I where X⁴ ismorpholin-4-yl.

In a still further embodiment, the compound has Formula I where R⁰ is

where X⁵ is O, CH₂, C(CH₃)₂ or CH₂CH₂; X⁶ and X⁷ are both hydrogen orboth methyl; and X⁸ is fluoro, chloro, bromo or iodo. Illustrativelyaccording to this embodiment X⁵ can be CH₂ or C(CH₃)₂ and/or each of X⁶and X⁷ can be methyl and/or X⁸ can be chloro.

In a still further embodiment, the compound has Formula I where R⁰ is

where X⁵ is O, CH₂, C(CH₃)₂ or CH₂CH₂; X⁶ and X⁷ are both hydrogen orboth methyl; and X⁸ is fluoro, chloro, bromo or iodo. Illustrativelyaccording to this embodiment X⁵ can be CH₂ or C(CH₃)₂ and/or each of X⁶and X⁷ can be methyl and/or X⁸ can be chloro.

In a still further embodiment, the compound has Formula I where X³ isfluoro and X⁴ is morpholin-4-yl.

In a still further embodiment, the compound has Formula I where X³ isfluoro and R⁰ is

where X⁵ is O, CH₂, C(CH₃)₂ or CH₂CH₂; X⁶ and X⁷ are both hydrogen orboth methyl; and X⁸ is fluoro, chloro, bromo or iodo. Illustrativelyaccording to this embodiment X⁵ can be CH₂ or C(CH₃)₂ and/or each of X⁶and X⁷ can be methyl and/or X⁸ can be chloro.

In a still further embodiment, the compound has Formula I where X⁴ ismorpholin-4-yl and R⁰ is

where X⁵ is O, CH₂, C(CH₃)₂ or CH₂CH₂; X⁶ and X⁷ are both hydrogen orboth methyl; and X⁸ is fluoro, chloro, bromo or iodo. Illustrativelyaccording to this embodiment X⁵ can be CH₂ or C(CH₃)₂ and/or each of X⁶and X⁷ can be methyl and/or X⁸ can be chloro.

In a still further embodiment, the compound has Formula I where X³ isfluoro, X⁴ is morpholin-4-yl and R⁰ is

where X⁵ is O, CH₂, C(CH₃)₂ or CH₂CH₂; X⁶ and X⁷ are both hydrogen orboth methyl; and X⁸ is fluoro, chloro, bromo or iodo. Illustrativelyaccording to this embodiment X⁵ can be CH₂ or C(CH₃)₂ and/or each of X⁶and X⁷ can be methyl and/or X⁸ can be chloro.

Compounds of Formula I may contain asymmetrically substituted carbonatoms in the R- or S-configuration; such compounds can be present asracemates or in an excess of one configuration over the other, forexample in an enantiomeric ratio of at least about 85:15. The compoundcan be substantially enantiomerically pure, for example having anenantiomeric ratio of at least about 95:5, or in some cases at leastabout 98:2 or at least about 99:1.

Compounds of Formula I may alternatively or additionally containcarbon-carbon double bonds or carbon-nitrogen double bonds in the Z- orE-configuration, the term “Z” denoting a configuration wherein thelarger substituents are on the same side of such a double bond and theterm “E” denoting a configuration wherein the larger substituents are onopposite sides of the double bond. The compound can alternatively bepresent as a mixture of Z- and E-isomers.

Compounds of Formula I may alternatively or additionally exist astautomers or equilibrium mixtures thereof wherein a proton shifts fromone atom to another. Examples of tautomers illustratively includeketo-enol, phenol-keto, oxime-nitroso, nitro-aci, imine-enamine and thelike.

Compounds of Formula I, and methods of preparation of such compounds,are disclosed in the above-cited '135 publication and/or in above-citedU.S. Patent Application Publication No. 2007/0072860, each of which isincorporated herein by reference in its entirety. Terms for substituentsused herein are defined exactly as in those publications.

In some embodiments, a compound of Formula I is present in thecomposition in its parent-compound (“free base”) form, alone or togetherwith a salt form of the compound.

Compounds of Formula I may form acid addition salts, basic additionsalts or zwitterions. Salts of compounds of Formula I can be preparedduring isolation or following purification of the compounds. Acidaddition salts are those derived from reaction of a compound of FormulaI with an acid. For example, salts including the acetate, adipate,alginate, bicarbonate, citrate, aspartate, benzoate, benzenesulfonate(besylate), bisulfate, butyrate, camphorate, camphorsulfonate,digluconate, formate, fumarate, glycerophosphate, glutamate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, lactobionate, lactate, maleate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate,pectinate, persulfate, phosphate, picrate, propionate, succinate,tartrate, thiocyanate, trichloroacetate, trifluoroacetate,para-toluenesulfonate and undecanoate salts of a compound of Formula Ican be used in a composition of the invention. Basic addition saltsincluding those derived from reaction of a compound with thebicarbonate, carbonate, hydroxide or phosphate of cations such aslithium, sodium, potassium, calcium and magnesium can likewise be used.

A compound of Formula I typically has more than one protonatablenitrogen atom and is consequently capable of forming acid addition saltswith more than one, for example about 1.2 to about 2, about 1.5 to about2 or about 1.8 to about 2, equivalents of acid per equivalent of thecompound.

ABT-263 (having Formula I where X³ is fluoro, X⁴ is morpholin-4-yl andR⁰ is

where X⁵ is —C(CH₃)₂—, X⁶ and X⁷ are both —H and X⁸ is chloro) canlikewise form acid addition salts, basic addition salts or zwitterions.Salts of ABT-263 can be prepared during isolation or followingpurification of the compound. Acid addition salts derived from reactionof ABT-263 with an acid include those listed above. Basic addition saltsincluding those listed above can likewise be used. ABT-263 has at leasttwo protonatable nitrogen atoms and is consequently capable of formingacid addition salts with more than one, for example about 1.2 to about2, about 1.5 to about 2 or about 1.8 to about 2, equivalents of acid perequivalent of the compound.

Illustratively in the case of ABT-263, bis-salts can be formedincluding, for example, bis-hydrochloride (bis-HCl) and bis-hydrobromide(bis-HBr) salts. These salts can alternatively be called ABT-263 diHCland ABT-263 diHBr.

For example, ABT-263 bis-HCl, which has a molecular weight of 1047.5g/mol and is represented by the formula

can be prepared by a variety of processes, for example a process thatcan be outlined as follows.

ABT-263 free base is prepared, illustratively as described in Example 1of the above-cited '135 publication, the entire disclosure of which isincorporated by reference herein. A suitable weight of ABT-263 free baseis dissolved in ethyl acetate. A solution of hydrochloric acid inethanol (for example about 4.3 kg HCl in 80 g EtOH) is added to theABT-263 solution in an amount providing at least 2 mol HCl per molABT-263 and sufficient EtOH (at least about 20 vol) for crystallizationof the resulting ABT-263 bis-HCl salt. The solution is heated to about45° C. with stiffing and seeds are added as a slurry in EtOH. Afterabout 6 hours, the resulting slurry is cooled to about 20° C. over about1 hour and is mixed at that temperature for about 36 hours. The slurryis filtered to recover a crystalline solid, which is an ethanol solvateof ABT-263 bis-HCl. Drying of this solid under vacuum and nitrogen withmild agitation for about 8 days yields white desolvated ABT-263 bis-HClcrystals. This material is suitable as active pharmaceutical ingredient(API) for preparation of an ABT-263 bis-HCl formulation of the presentinvention.

The term “free base” is used for convenience herein to refer to theparent compound, while recognizing that the parent compound is, strictlyspeaking, zwitterionic and thus does not always behave as a true base.ABT-263 bis-HCl can be prepared by any process that comprises reactingABT-263 free base with 2 moles of hydrochloric acid (HCl) in a suitablemedium.

As indicated above, ABT-263 free base can be prepared by a process asdescribed in Example 1 of the above-cited '135 publication. The productof this process is an amorphous, glassy solid. A powder can be preparedfrom this product, for example by freeze-drying or precipitationtechniques. Such a powder can be used as API in preparing a compositionof the present invention; however, it will generally be found preferableto use a crystalline form of ABT-263 free base as API. Such crystallineforms include solvates and solvent-free crystalline forms.

Solvates of ABT-263 free base can be prepared as described below. Thestarting product can be any solid-state form of ABT-263 free base,including the amorphous form prepared according to the '135 publication.

A measured amount of ABT-263 free base (as indicated, any solid-stateform can be used) is suspended in any of a number of solvents or solventmixtures, including without limitation 2-propanol, 1-propanol, ethylacetate/ethanol 1:3 v/v, methyl acetate/hexanes 1:1 v/v, chloroform,methanol, 1,4-dioxane/hexanes 1:2 v/v, toluene and benzene. Theresulting suspension is agitated at ambient temperature, while protectedfrom light. After a period of time sufficient to permit solvation ofABT-263 free base in each case, crystals are harvested by filtercentrifugation. The resulting solvates can be characterized by powderX-ray diffraction (PXRD), for example using a G3000 diffractometer (InelCorp., Artenay, France) equipped with a curved position-sensitivedetector and parallel-beam optics. The diffractometer is operated with acopper anode tube (1.5 kW fine focus) at 40 kV and 30 mA. Anincident-beam germanium monochromator provides monochromatic radiation.The diffractometer is calibrated using an attenuated direct beam atone-degree intervals. Calibration is checked using a silicon powder lineposition reference standard (NIST 640c). The instrument iscomputer-controlled using Symphonix software (Inel Corp., Artenay,France) and the data are analyzed using Jade software (version 6.5,Materials Data, Inc., Livermore, Calif.). The sample is loaded onto analuminum sample holder and leveled with a glass slide.

Desolvation of an ethyl acetate/ethanol solvate, for example byair-drying, provides a solvent-free crystalline form of ABT-263 freebase. PXRD peaks for Form I ABT-263 free base are listed in Table 1. APXRD pattern having peaks substantially as indicated therein can be usedto identify crystalline ABT-263 free base, more particularly Form IABT-263 free base. The phrase “substantially as indicated” in thepresent context means having peaks that are not shifted more than about0.2° 2θ from the indicated position.

TABLE 1 PXRD peak listing: solvent-free crystal polymorph Form I ABT-263free base Peak Position (° 2θ) 6.21 6.72 9.66 10.92 11.34 12.17 14.2816.40 16.95 17.81 18.03 18.47 19.32 20.10 21.87

Desolvation of most solvates, including 1-propanol, 2-propanol,methanol, benzene, toluene, dioxane/hexanes, methyl acetate/hexanes andchloroform solvates, provides a solvent-free crystalline form of ABT-263free base that is shown by PXRD to be identical to the crystalline formproduced by desolvation of the ethyl acetate/ethanol solvate.

Desolvation of pyridine and anisole solvates provides a solvent-freecrystalline form of ABT-263 free base that is shown by PXRD to bedifferent from the form produced by desolvation of the ethylacetate/ethanol solvate. The crystalline form derived from desolvationof the pyridine or anisole solvate is designated Form II. A PXRD scan ofForm II ABT-263 free base is shown in FIG. 2. PXRD peaks for Form IIABT-263 free base are listed in Table 2. A PXRD pattern having peakssubstantially as indicated therein can be used to identify crystallineABT-263 free base, more particularly Form II ABT-263 free base.

TABLE 2 PXRD peak listing: solvent-free crystal polymorph Form IIABT-263 free base Peak Position (° 2θ) 5.79 8.60 9.34 10.79 11.36 11.5912.76 13.23 13.73 14.01 14.72 15.00 16.28 17.07 17.48 18.75 19.34 19.7120.56 21.35

PXRD peaks especially diagnostic for Form I ABT-263 free base, inparticular for distinguishing Form I from Form II, include the peaks at6.21, 6.72, 12.17, 18.03 and 20.10° 20, in each case ±0.2° 2θ. In oneembodiment, Form I ABT-263 free base is characterized at least by a peakat any one or more of these positions. In another embodiment, Form IABT-263 free base is characterized at least by a peak at each of thesepositions. In yet another embodiment, Form I ABT-263 free base ischaracterized by a peak at each of the positions shown in Table 1.

PXRD peaks especially diagnostic for Form II ABT-263 free base, inparticular for distinguishing Form II from Form I, include the peaks at5.79, 8.60, 12.76, 15.00 and 20.56° 2θ, in each case ±0.2° 2θ. In oneembodiment, Form II ABT-263 free base is characterized at least by apeak at any one or more of these positions. In another embodiment, FormII ABT-263 free base is characterized at least by a peak at each ofthese positions. In yet another embodiment, Form II ABT-263 free base ischaracterized by a peak at each of the positions shown in Table 2.

Any of the crystalline forms of ABT-263 free base, including solvatedforms, can be useful as API for preparation of a capsule of the presentinvention. However, solvent-free forms such as Form I and Form II aregenerally preferred for this purpose.

Without being bound by theory, it is believed that the therapeuticefficacy of compounds of Formula I is due at least in part to theirability to bind to a Bcl-2 family protein such as Bcl-2, Bcl-X_(L) orBcl-w in a way that inhibits the anti-apoptotic action of the protein,for example by occupying the BH3 binding groove of the protein. It willgenerally be found desirable to select a compound having high bindingaffinity for a Bcl-2 family protein, for example a K_(i) not greaterthan about 5 nM, preferably not greater than about 1 nM.

A composition as provided herein comprising any specific compounddisclosed in the '135 publication is expressly contemplated as anembodiment of the present invention.

In a more particular embodiment, the composition comprises ABT-263 or asalt thereof. In a still more particular embodiment, the compositioncomprises ABT-263 free base or a salt, for example a bis-salt, thereof.In an even more particular embodiment, the composition comprises ABT-263free base or ABT-263 bis-HCl.

Amounts, concentrations and dosages of a compound of Formula I or a saltthereof, for example of ABT-263 free base or ABT-263 bis-HCl, areexpressed herein as free base equivalent, unless the context demandsotherwise. Illustratively, in the case of ABT-263 bis-HCl, 1 mg freebase equivalent translates to about 1.075 mg of the salt. Unlessotherwise indicated, concentrations expressed as percentages herein areby weight.

A composition of the present invention contains a compound of Formula Ior a salt thereof, for example ABT-263 free base or ABT-263 bis-HCl, ina free base equivalent amount of at least about 2.5% by weight. Anactive ingredient concentration in a liquid composition indicated hereinto be 25 mg/l (a weight/volume concentration) will be understood to be“about 2.5% by weight” and at least in that regard within the scope ofthe present invention. An upper limit of concentration of a compound ofFormula I or a salt thereof, for example ABT-263 free base or ABT-263bis-HCl, in a composition is dictated by physical constraints such asdrug solubility in the case of liquid solution compositions and byamounts of excipient ingredients required, e.g., for acceptablebioavailability, in the case of solid compositions, but is unlikely toexceed about 50% by weight.

In various embodiments, the free base equivalent concentration of thesole or first active ingredient in the composition is at least about 3%,at least about 4%, at least about 5% or at least about 10%, by weight,or at least about 30 mg/l, at least about 40 mg/l, at least about 50mg/l or at least about 100 mg/l.

The sole or first active ingredient is present in the composition in anamount that can be therapeutically effective when the composition isadministered to a subject in need thereof according to an appropriateregimen. Typically, a unit dose (the amount administered at a singletime), which can be administered at an appropriate frequency, e.g.,twice daily to once weekly, is about 10 to about 1,000 mg free baseequivalent, depending on the compound in question. Where frequency ofadministration is once daily (q.d.), unit dose and daily dose are thesame. Illustratively, for example where the drug is ABT-263, the unitdose is typically about 25 to about 1,000 mg, more typically about 50 toabout 500 mg, for example about 50, about 100, about 150, about 200,about 250, about 300, about 350, about 400, about 450 or about 500 mg.Where the composition is provided as discrete dosage forms such ascapsules or tablets, the unit dose can generally be delivered in one toa small plurality, most typically 1 to about 10, such dosage forms. Thehigher the unit dose, the more desirable it becomes to select aformulation with a relatively high concentration of the drug therein.

Necessarily where the sole or first active ingredient is in solution ina liquid carrier, and optionally where the sole or first activeingredient is in solid-state form as defined herein, the compositionfurther comprises an antioxidant.

An “antioxidant” or compound having “antioxidant” properties is achemical compound that prevents, inhibits, reduces or retards oxidationof another chemical or itself. Antioxidants can improve stability andshelf-life of a lipid formulation as described herein by, for example,preventing, inhibiting, reducing or retarding oxidation of the compoundof Formula I in the formulation.

Enhancement of stability or shelf-life can be evaluated, for example, bymonitoring rate of appearance or build-up of sulfoxides in theformulation. Sulfoxides in total can be monitored by repeated samplingand analysis; alternatively samples can be analyzed more specificallyfor the sulfoxide degradation product of the compound of Formula I,i.e., the compound having the formula

where X³, X⁴ and R⁰ are as indicated above; or the sulfoxide degradationproduct of ABT-263, having the formula

Reference herein to the sulfoxide degradation product will be understoodto include both diastereomers at the sulfur atom stereocenter in thesulfoxide group.

An “antioxidant effective amount” of an antioxidant herein is an amountthat provides

-   -   (a) a substantial reduction (for example a reduction of at least        about 25%, at least about 50%, at least about 75%, at least        about 80%, at least about 85% or at least about 90%) in the        formation or accumulation of a degradation product, for example        the sulfoxide degradation product above, and/or    -   (b) a substantial increase (for example at least about 30, at        least about 60, at least about 90 or at least about 180 days) in        the time taken for the degradation product to reach a threshold        level,        in a formulation containing the antioxidant, by comparison with        an otherwise similar formulation containing no antioxidant. A        storage-stability study to determine degree of (a) reduction in        formation or accumulation of the degradation product or (b)        increase in time taken for a degradation product to reach a        threshold level in the formulation can be conducted at any        appropriate temperature or range of temperatures.        Illustratively, a study at about 5° C. can be indicative of        storage stability under refrigerated conditions, a study at        about 20-25° C. can be indicative of storage stability under        typical ambient conditions, and a study at about 30° C. or        higher temperature can be useful in an accelerated-aging study.        Any appropriate threshold level of the degradation product can        be selected as an end-point, for example in the range from about        0.2% to about 2% of the initial amount of the compound of        Formula I present.

In various illustrative embodiments, the antioxidant is included in anamount effective to hold oxidative degradation of the drug

(a) below about 1% for at least about 3 months;

(b) below about 1% for at least about 6 months;

(c) below about 1% for at least about 1 year;

(d) below about 0.5% for at least about 3 months;

(e) below about 0.5% for at least about 6 months; or

(f) below about 0.5% for at least about 1 year;

in the formulation when stored under ambient conditions (e.g., about20-25° C.) in a sealed container opaque to ultraviolet light, asmeasured for example by amount of the sulfoxide degradation productpresent at the end of the recited storage period.

Antioxidants used in pharmaceutical compositions are most typicallyagents that inhibit generation of oxidative species such as triplet orsinglet oxygen, superoxides, peroxide and free hydroxyl radicals, oragents that scavenge such oxidative species as they are generated.Examples of commonly used antioxidants of these classes includebutylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), retinylpalmitate, tocopherol, propyl gallate, ascorbic acid and ascorbylpalmitate. The present inventors have found, however, that at least somecommonly used antioxidants are ineffective to protect ABT-263 fromexcessive sulfoxide formation in encapsulated liquid formulations asdescribed herein.

For example, BHA, added at 0.2% by weight to a 15% by weight solution ofABT-263 free base in a medium referred to herein as “IPT-253” (20%Imwitor 742™, 50% Phosal 53 MCT™, 30% Tween™ 80), has been found to haveno effect on sulfoxide formation in a 4-week stability study at 40° C.without nitrogen purging of headspace, as shown in Table 3. A fullreport of this study is found in Example 7 herein.

TABLE 3 Effect of 0.2% BHA on ABT-263 sulfoxide formation in IPT-253solution Time % Total sulfoxides (weeks) No antioxidant 0.2% BHA 0 notdetectable 0.06 1 0.26 0.29 2 0.47 0.49 3 0.56 0.58 4 0.67 0.68

Antioxidants that, by contrast, have been found effective areheavier-chalcogen antioxidants (HCAs) that are believed, without beingbound by theory, to function primarily as competitive substrates, i.e.,as “sacrificial” antioxidants, which are preferentially attacked byoxidative species thereby protecting the drug from excessivedegradation.

In some embodiments, the HCA comprises one or more antioxidant compoundsof Formula II

where

n is 0, 1 or 2;

Y¹ is S or Se;

Y² is NHR¹, OH or H, where R¹ is alkyl or alkylcarbonyl;

Y³ is COOR² or CH₂OH, where R² is H or alkyl; and

R³ is H or alkyl;

where alkyl groups are independently optionally substituted with one ofmore substituents independently selected from the group consisting ofcarboxyl, alkylcarbonyl, alkoxycarbonyl, amino and alkylcarbonylamino; apharmaceutically acceptable salt thereof; or, where Y¹ is S and R³ is H,an —S—S— dimer thereof or pharmaceutically acceptable salt of suchdimer.

In other embodiments, the HCA is an antioxidant compound of Formula III:

where

-   -   Y is S, Se or S—S; and    -   R⁴ and R⁵ are independently selected from H, alkyl and        (CH₂)_(n)R⁶ where n is 0-10 and R⁶ is arylcarbonyl,        alkylcarbonyl, alkoxycarbonyl, carboxyl or CHR⁷R⁸-substituted        alkyl, where R⁷ and R⁸ are independently CO₂R⁹, CH₂OH, hydrogen        or NHR¹⁰, where R⁹ is H, alkyl, substituted alkyl or arylalkyl        and R¹⁰ is hydrogen, alkyl, alkylcarbonyl or alkoxycarbonyl.

An “alkyl” substituent or an “alkyl” or “alkoxy” group forming part of asubstituent according to Formula II or Formula III is one having 1 toabout 18 carbon atoms and can consist of a straight or branched chain.

An “aryl” group forming part of a substituent according to Formula IIIis a phenyl group, unsubstituted or substituted with one or morehydroxy, alkoxy or alkyl groups.

In some embodiments, R¹ in Formula II is C₁₋₄ alkyl (e.g., methyl orethyl) or (C₁₋₄ alkyl)carbonyl (e.g., acetyl).

In some embodiments, R² in Formula II is H or C₁₋₁₈ alkyl, for examplemethyl, ethyl, propyl (e.g., n-propyl or isopropyl), butyl (e.g.,n-butyl, isobutyl or t-butyl), octyl (e.g., n-octyl or 2-ethylhexyl),dodecyl (e.g., lauryl), tridecyl, tetradecyl, hexadecyl or octadecyl(e.g., stearyl).

R³ is typically H or C₁₋₄ alkyl (e.g., methyl or ethyl).

The HCA can be, for example, a natural or synthetic amino acid or aderivative thereof such as an alkyl ester or N-acyl derivative, or asalt of such amino acid or derivative. Where the amino acid orderivative thereof is derived from a natural source it is typically inthe L-configuration; however it is understood that D-isomers andD,L-isomer mixtures can be substituted if necessary.

Non-limiting examples of HCAs useful herein includeβ-alkylmercaptoketones, cysteine, cystine, homocysteine, methionine,thiodiglycolic acid, thiodipropionic acid, thioglycerol, selenocysteine,selenomethionine and salts, esters, amides and thioethers thereof; andcombinations thereof. More particularly, one or more HCAs can beselected from N-acetylcysteine, N-acetylcysteine butyl ester,N-acetylcysteine dodecyl ester, N-acetyl-cysteine ethyl ester,N-acetylcysteine methyl ester, N-acetylcysteine octyl ester,N-acetyl-cysteine propyl ester, N-acetylcysteine stearyl ester,N-acetylcysteine tetradecyl ester, N-acetylcysteine tridecyl ester,N-acetylmethionine, N-acetylmethionine butyl ester, N-acetylmethioninedodecyl ester, N-acetylmethionine ethyl ester, N-acetylmethionine methylester, N-acetylmethionine octyl ester, N-acetylmethionine propyl ester,N-acetylmethionine stearyl ester, N-acetylmethionine tetradecyl ester,N-acetylmethionine tridecyl ester, N-acetyl-selenocysteine,N-acetylselenocysteine butyl ester, N-acetylselenocysteine dodecylester, N-acetylselenocysteine ethyl ester, N-acetylselenocysteine methylester, N-acetylseleno-cysteine octyl ester, N-acetylselenocysteinepropyl ester, N-acetylselenocysteine stearyl ester,N-acetylselenocysteine tetradecyl ester, N-acetylselenocysteine tridecylester, N-acetylseleno-methionine, N-acetylselenomethionine butyl ester,N-acetylselenomethionine dodecyl ester, N-acetylselenomethionine ethylester, N-acetylselenomethionine methyl ester, N-acetyl-selenomethionineoctyl ester, N-acetylselenomethionine propyl ester,N-acetylseleno-methionine stearyl ester, N-acetylselenomethioninetetradecyl ester, N-acetylseleno-methionine tridecyl ester, cysteine,cysteine butyl ester, cysteine dodecyl ester, cysteine ethyl ester,cysteine methyl ester, cysteine octyl ester, cysteine propyl ester,cysteine stearyl ester, cysteine tetradecyl ester, cysteine tridecylester, cystine, cystine dibutyl ester, cystine di(dodecyl) ester,cystine diethyl ester, cystine dimethyl ester, cystine dioctyl ester,cystine dipropyl ester, cystine distearyl ester, cystine di(tetradecyl)ester, cystine di(tridecyl) ester, N,N-diacetylcystine,N,N-diacetylcystine dibutyl ester, N,N-diacetylcystine diethyl ester,N,N-diacetylcystine di(dodecyl) ester, N,N-diacetylcystine dimethylester, N,N-diacetylcystine dioctyl ester, N,N-diacetylcystine dipropylester, N,N-diacetylcystine distearyl ester, N,N-diacetylcystinedi(tetradecyl) ester, N,N-diacetylcystine di(tridecyl) ester, dibutylthiodiglycolate, dibutyl thiodipropionate, di(dodecyl) thiodiglycolate,di(dodecyl) thiodipropionate, diethyl thiodiglycolate, diethylthiodipropionate, dimethyl thiodiglycolate, dimethyl thiodipropionate,dioctyl thiodiglycolate, dioctyl thiodipropionate, dipropylthiodiglycolate, dipropyl thiodipropionate, distearyl thiodiglycolate,distearyl thiodipropionate, di(tetradecyl) thiodiglycolate,di(tetradecyl) thiodipropionate, homocysteine, homocysteine butyl ester,homocysteine dodecyl ester, homocysteine ethyl ester, homocysteinemethyl ester, homocysteine octyl ester, homocysteine propyl ester,homocysteine stearyl ester, homocysteine tetradecyl ester, homocysteinetridecyl ester, methionine, methionine butyl ester, methionine dodecylester, methionine ethyl ester, methionine methyl ester, methionine octylester, methionine propyl ester, methionine stearyl ester, methioninetetradecyl ester, methionine tridecyl ester, S-methylcysteine,S-methyl-cysteine butyl ester, S-methylcysteine dodecyl ester,S-methylcysteine ethyl ester, S-methyl-cysteine methyl ester,S-methylcysteine octyl ester, S-methylcysteine propyl ester,S-methyl-cysteine stearyl ester, S-methylcysteine tetradecyl ester,S-methylcysteine tridecyl ester, selenocysteine, selenocysteine butylester, selenocysteine dodecyl ester, selenocysteine ethyl ester,selenocysteine methyl ester, selenocysteine octyl ester, selenocysteinepropyl ester, selenocysteine stearyl ester, selenocysteine tetradecylester, selenocysteine tridecyl ester, selenomethionine, selenomethioninebutyl ester, selenomethionine dodecyl ester, seleno-methionine ethylester, selenomethionine methyl ester, selenomethionine octyl ester,seleno-methionine propyl ester, selenomethionine stearyl ester,selenomethionine tetradecyl ester, selenomethionine tridecyl ester,thiodiglycolic acid, thiodipropionic acid, thioglycerol, isomers andmixtures of isomers thereof, and salts thereof.

In some embodiments, the HCA selected is a sulfur-containingantioxidant.

Salts of HCA compounds can be acid addition salts such as the acetate,adipate, alginate, bicarbonate, citrate, aspartate, benzoate,benzenesulfonate (besylate), bisulfate, butyrate, camphorate,camphorsulfonate, digluconate, formate, fumarate, glycerophosphate,glutamate, hemisulfate, heptanoate, hexanoate, hydrochloride,hydrobromide, hydroiodide, lactobionate, lactate, maleate,mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nicotinate,oxalate, pamoate, pectinate, persulfate, phosphate, picrate, propionate,succinate, tartrate, thiocyanate, trichloroacetate, trifluoroacetate,para-toluenesulfonate and undecanoate salts. In a particular embodiment,the hydrochloride salt of one of the compounds individually mentionedabove is present in the composition in an antioxidant effective amount.

Without being bound by theory, it is generally believed that HCAs suchas those exemplified above protect the active compound by beingthemselves more readily oxidizable and, therefore, being oxidizedpreferentially over the drug compound. In general, for this mode ofoperation to provide an acceptable degree of protection for the drugcompound, an antioxidant of Formula II or Formula III must be present ina substantial amount, for example in a molar ratio to the drug compoundof at least about 1:10. In some embodiments, the molar ratio ofantioxidant to the drug compound is about 1:10 to about 2:1, for exampleabout 1:5 to about 1.5:1. Best results will sometimes be obtained whenthe molar ratio is approximately 1:1, i.e., about 8:10 to about 10:8.

This typical requirement for a relatively high antioxidant concentrationin the formulation places constraints both on the selection ofantioxidant and on the selection of other formulation components,particularly in liquid solution compositions of the invention. For suchcompositions, a carrier system must be selected that is capable ofdissolving not only the active agent but also the antioxidant, in anantioxidant effective amount. One of skill in the art can select asuitable lipid carrier, which can comprise a single lipid material or amixture of two or more such materials, by routine solubility testingbased on the disclosure herein.

Notwithstanding the antioxidant efficacy of HCAs of Formula II orFormula III, the present inventors have found that, at molar ratios ofapproximately 1:1, such antioxidants have a tendency to result insolutions that become cloudy upon storage, when ABT-263 is used in theform of its free base. For solutions containing ABT-263 in the form ofits bis-HCl salt, this tendency is absent or at least less marked.

However, in yet another unexpected discovery, ABT-263 free base has beenfound to be less susceptible to sulfoxide formation than ABT-263 bis-HClwhen formulated in lipid solution (but in the absence of antioxidant),as shown in Table 6 (see Example 3 hereinbelow). The solvent system inSolution A is Phosal 53 MCT™/ethanol, 9:1 v/v; and in Solution B isLabrafil M 1944 CS™/oleic acid/polysorbate 80, 30%/40%/30% by weight.(Labrafil M 1944 CS™ of Gattefossé contains polyoxyethylene glycerylmonooleate.) The three-week study was conducted at 40° C. withoutnitrogen purging of headspace.

To take advantage of the unexpected finding that ABT-263 is lesssusceptible to sulfoxide formation in its free base than salt form, thepresent inventors have turned to a different class of sulfur-containingantioxidants, namely inorganic antioxidants of the sulfite, bisulfite,metabisulfite and thiosulfate classes. To complicate matters, theseantioxidants are poorly lipid-soluble and must be introduced to thecarrier or drug-carrier system in aqueous solution. Presence of waterpromotes sulfoxide formation in ABT-263 solutions, the very effect thatis sought to be minimized. To restrict the amount of added water, poorlylipid-soluble antioxidants are, in one embodiment of the presentinvention, added at much lower concentrations than those providing molarequivalence to the concentration of ABT-263.

Where a poorly lipid-soluble antioxidant such as a sulfite, bisulfite,metabisulfite or thiosulfate antioxidant is used, it is accompanied inthe composition by water in an amount not exceeding about 1% by weight,for example about 0.2% to about 0.8% by weight. The amount of suchantioxidant that can be introduced in such a small amount of watertypically does not exceed about 0.2% by weight, and is for example anamount of about 0.02% to about 0.2%, or about 0.05% to about 0.15%, byweight, of the composition.

To minimize the amount of water added to the formulation, it isdesirable to provide the antioxidant in the form of a relativelyconcentrated aqueous stock solution, for example having at least about10% by weight antioxidant. However, it has been found that where anexcessively concentrated stock solution (e.g., about 20% or higher) isused, this can result in undesirable precipitation of solids in theformulation. Suitable concentrations of antioxidant in the stocksolution are typically about 10% to about 18%, illustratively about 15%,by weight.

Sodium and potassium salts of sulfites, bisulfites, metabisulfites andthiosulfates are useful antioxidants according to the presentembodiment; more particularly sodium and potassium metabisulfites.

To further minimize sulfoxide formation, a chelating agent such as EDTAor a salt thereof (e.g., disodium EDTA or calcium disodium EDTA) isoptionally added, for example in an amount of about 0.002% to about0.02% by weight of the composition. EDTA can be added as an aqueousstock solution in the same manner as the antioxidant. The antioxidantand EDTA can, if desired, be added as components of the same stocksolution. Chelating agents sequester metal ions that can promoteoxidative degradation.

Surprisingly at the very low antioxidant concentrations contemplatedherein (typically the molar ratio of poorly lipid-soluble antioxidant toABT-263 according to the present embodiment is no greater than about1:20), sulfoxide formation has been found to remain within acceptablelimits, as illustrated in Example 12 herein.

Sulfoxide formation can be further minimized by selecting formulationingredients having low peroxide value. Peroxide value is a wellestablished property of pharmaceutical excipients and is generallyexpressed (as herein) in units corresponding to milliequivalents ofperoxides per kilogram of excipient (meq/kg). Some excipients inherentlyhave low peroxide value, but others, for example those havingunsaturated fatty acid such as oleyl moieties and/or polyoxyethylenechains, can be sources of peroxides. In the case of polysorbate 80, forexample, it is preferable to select a source of polysorbate 80 having aperoxide value not greater than about 5, for example not greater thanabout 2. Suitable sources include Crillet 4HP™ and Super-Refined Tween™80, both available from Croda.

First Composition Embodiment

A composition of the first embodiment set forth hereinabove comprises(a) a compound of Formula I or a pharmaceutically acceptable saltthereof, in a free base equivalent amount of at least about 2.5% byweight of the composition; (b) a pharmaceutically acceptable HCA; and(c) a substantially non-aqueous pharmaceutically acceptable carrier thatcomprises one or more lipids; wherein said compound and the antioxidantare in solution in the carrier.

The term “drug-carrier system” as used in description of compositions ofthe present embodiment comprises a carrier having at least one drughomogeneously distributed therein. In such compositions the drug (acompound of Formula I or a salt thereof) and HCA are in solution in thecarrier, and, in some of these compositions, the drug-carrier systemconstitutes essentially the entire composition. In other compositions,the drug-carrier system is encapsulated within a capsule shell that issuitable for oral administration; in such embodiments the compositioncomprises the drug-carrier system and the capsule shell.

A drug-carrier system of the present embodiment is typically liquid, butin some compositions the carrier and/or the drug-carrier system can besolid or semi-solid. For example, a drug-carrier system canillustratively be prepared by dissolving the drug and HCA in a carrierat a temperature above the melting or flow point of the carrier, andcooling the resulting solution to a temperature below the melting orflow point to provide a solid drug-carrier system. The drug-carriersystem can optionally comprise a solid or semi-solid substrate havingthe drug solution adsorbed therein or thereon. Examples of suchsubstrates include particulate diluents such as lactose, starches,silicon dioxide, etc., and polymers such as polyacrylates, highmolecular weight PEGs, or cellulose derivatives, e.g.,hydroxypropylmethylcellulose (HPMC). Where a solid solution is desired,a high melting point ingredient such as a wax can be included. A soliddrug-carrier system can optionally be encapsulated or, if desired,delivered in tablet form. The drug-carrier system can, in someembodiments, be adsorbed on, or impregnated into, a drug deliverydevice.

In a composition of the present embodiment, the drug is “in solution” inthe carrier. This will be understood to mean that substantially all ofthe drug is in solution, i.e., no substantial portion, for example nomore than about 2%, or no more than about 1%, of the drug is in solid(e.g., crystalline) form, whether dispersed, for example in the form ofa suspension, or not. In practical terms, this means that the drug mustnormally be formulated at a concentration below its limit of solubilityin the carrier. It will be understood that the limit of solubility canbe temperature-dependent, thus selection of a suitable concentrationshould take into account the range of temperatures to which thecomposition is likely to be exposed in normal storage, transport anduse.

Not only the drug, but also the HCA, is “in solution” as defined abovein the carrier. Where the HCA is poorly lipid-soluble and has to beintroduced to the carrier or drug-carrier system in aqueous solution, asurfactant, more particularly a non-phospholipid surfactant, may benecessary to avoid phase separation.

The carrier according to the present embodiment is “substantiallynon-aqueous”, i.e., having no water, or having an amount of water thatis small enough to be, in practical terms, essentially non-deleteriousto performance or properties of the composition. Typically, the carriercomprises zero to less than about 5% by weight water. It will beunderstood that certain ingredients useful herein can bind small amountsof water on or within their molecules or supramolecular structures; suchbound water if present does not affect the “substantially non-aqueous”character of the carrier as defined herein. Furthermore, as indicatedabove, use of a poorly lipid-soluble antioxidant requires that a smallamount of water (not more than about 1% by weight of the drug-carriersystem) be added; again, this does not affect the “substantiallynon-aqueous” character of the carrier as defined herein.

In some compositions, the carrier comprises one or more glyceridematerials. Suitable glyceride materials include, without limitation,medium to long chain mono-, di- and triglycerides. The term “mediumchain” herein refers to hydrocarbyl chains individually having no lessthan about 6 and less than about 12 carbon atoms, including for exampleC₈ to C₁₀ chains. Thus glyceride materials comprising caprylyl andcapryl chains, e.g., caprylic/capric mono-, di- and/or triglycerides,are examples of “medium chain” glyceride materials herein. The term“long chain” herein refers to hydrocarbyl chains individually having atleast about 12, for example about 12 to about 18, carbon atoms,including for example lauryl, myristyl, cetyl, stearyl, oleyl, linoleyland linolenyl chains. Medium to long chain hydrocarbyl groups in theglyceride materials can be saturated, mono- or polyunsaturated.

In one embodiment the carrier comprises a medium chain and/or a longchain triglyceride material. A suitable example of a medium chaintriglyceride material is a caprylic/capric triglyceride product such asCaptex 355 EP™ of Abitec Corp. and products substantially equivalentthereto. Suitable examples of long chain triglycerides include anypharmaceutically acceptable vegetable oil, for example canola, coconut,corn, cottonseed, flaxseed, olive, palm, peanut, safflower, sesame, soyand sunflower oils, and mixtures of such oils. Oils of animal,particularly marine animal, origin can also be used, including forexample fish oil.

A carrier system that has been found particularly useful in solubilizingboth (a) a therapeutically effective amount of a compound of Formula Iand (b) an antioxidant effective amount of an HCA, comprises twoessential components: a phospholipid, and a pharmaceutically acceptablesolubilizing agent for the phospholipid. It will be understood thatreference in the singular to a (or the) phospholipid, solubilizing agentor other formulation ingredient herein includes the plural; thuscombinations, for example mixtures, of more than one phospholipid, ormore than one solubilizing agent, are expressly contemplated herein. Thesolubilizing agent, or the combination of solubilizing agent andphospholipid, also solubilizes the drug and the antioxidant, althoughother carrier ingredients, such as a surfactant or an alcohol such asethanol, optionally present in the carrier can in some circumstancesprovide enhanced solubilization of the drug and antioxidant.

Any pharmaceutically acceptable phospholipid or mixture of phospholipidscan be used. In general such phospholipids are phosphoric acid estersthat yield on hydrolysis phosphoric acid, fatty acid(s), an alcohol anda nitrogenous base. Pharmaceutically acceptable phospholipids caninclude without limitation phosphatidylcholines, phosphatidylserines andphosphatidylethanolamines. In one embodiment the composition comprisesphosphatidylcholine, derived for example from natural lecithin. Anysource of lecithin can be used, including animal sources such as eggyolk, but plant sources are generally preferred. Soy is a particularlyrich source of lecithin that can provide phosphatidylcholine for use inthe present invention.

Illustratively, a suitable amount of phospholipid is about 15% to about75%, for example about 30% to about 60%, by weight of the carrier,although greater and lesser amounts can be useful in particularsituations.

Ingredients useful as components of the solubilizing agent are notparticularly limited and will depend to some extent on the particulardrug and HCA and the desired concentration of each and of phospholipid.In one embodiment, the solubilizing agent comprises one or more glycols,one or more glycolides and/or one or more glyceride materials.

Glycols are generally suitable only for non-encapsulated formulations orwhere a soft capsule shell is to be used, and tend to be incompatiblewith hard shells such as hard gelatin shells. Suitable glycols includepropylene glycol and polyethylene glycols (PEGs) having molecular weightof about 200 to about 1,000 g/mol, e.g., PEG-400, which has an averagemolecular weight of about 400 g/mol. Such glycols can provide relativelyhigh solubility of the drug; however the potential for oxidativedegradation of the drug can be increased when in solution in a carriercomprising such glycols, for example because of the tendency of glycolsto produce superoxides, peroxides and/or free hydroxyl radicals. Thehigher the glycol content of the carrier, the greater may be thetendency for degradation of a chemically unstable drug. In oneembodiment, therefore, one or more glycols are present in a total glycolamount of at least about 1% but less than about 50%, for example lessthan about 30%, less than about 20%, less than about 15% or less thanabout 10% by weight of the carrier. In another embodiment, the carriercomprises substantially no glycol.

Glycolides are glycols such as propylene glycol or PEG esterified withone or more organic acids, for example medium- to long-chain fattyacids. Suitable examples include propylene glycol monocaprylate,propylene glycol monolaurate and propylene glycol dilaurate productssuch as, for example. Capmul PG8™, Capmul PG12™ and Capmul PG-2L™respectively of Abitec Corp. and products substantially equivalentthereto.

Suitable glyceride materials for use together with a phospholipidinclude, without limitation, those mentioned above. Where one or moreglyceride materials are present as a major component of the solubilizingagent, a suitable total amount of glycerides is an amount effective tosolubilize the phospholipid and, in combination with other components ofthe carrier, effective to maintain the drug and antioxidant in solution.For example, glyceride materials such as medium chain and/or long chainmono-, di- and triglycerides, more typically medium-chain mono-, di- andtriglycerides, can be present in a total glyceride amount of about 5% toabout 70%, for example about 15% to about 60% or about 25% to about 50%,by weight of the carrier, although greater and lesser amounts can beuseful in particular situations. In one embodiment, the encapsulatedliquid comprises about 7% to about 30%, for example about 10% to about25%, by weight medium-chain triglycerides and about 7% to about 30%, forexample about 10% to about 25%, by weight medium-chain mono- anddiglycerides.

Additional solubilizing agents that are other than glycols, glycolidesor glyceride materials can be included if desired. Such agents, forexample N-substituted amide solvents such as dimethylformamide (DMF) andN,N-dimethylacetamide (DMA), can, in specific cases, assist in raisingthe limit of solubility of the drug in the carrier, thereby permittingincreased drug loading. However, the carriers useful herein generallyprovide adequate solubility of small-molecule drugs of interest hereinwithout such additional agents.

Even when a sufficient amount of a glycol, glycolide or glyceridematerial is present to solubilize the phospholipid, the resultingcarrier solution and/or the drug-carrier system may be rather viscousand difficult or inconvenient to handle. In such cases it may be founddesirable to include in the carrier a viscosity reducing agent in anamount effective to provide acceptably low viscosity. An example of suchan agent is an alcohol, more particularly ethanol, which is preferablyintroduced in a form that is substantially free of water, for example99% ethanol, dehydrated alcohol USP or absolute ethanol. Excessivelyhigh concentrations of ethanol should, however, generally be avoided.This is particularly true where, for example, the drug-carrier system isto be administered in a gelatin capsule, because of the tendency of highethanol concentrations to result in mechanical failure of the capsule.In general, suitable amounts of ethanol are 0% to about 25%, for exampleabout 1% to about 20% or about 3% to about 15%, by weight of thecarrier. Glycols such as propylene glycol or PEG and medium-chain mono-and diglycerides (for example caprylic/capric mono- and diglycerides)can also be helpful to lower viscosity; where the drug-carrier system isto be encapsulated in a hard capsule such as a hard gelatin capsule,medium-chain mono- and diglycerides are particularly useful in thisregard.

Optionally, the carrier further comprises a pharmaceutically acceptablenon-phospholipid surfactant. One of skill in the art will be able toselect a suitable surfactant for use in a composition of the presentembodiment, based on information herein. Such a surfactant can servevarious functions, including for example enhancing dispersion of theencapsulated liquid upon release from the capsule in the aqueousenvironment of the gastrointestinal tract. Thus in one embodiment thenon-phospholipid surfactant is a dispersing and/or emulsifying agentthat enhances dispersion and/or emulsification of the capsule contentsin real or simulated gastrointestinal fluid. Illustratively, asurfactant such as a polysorbate (polyoxyethylene sorbitan ester), e.g.,polysorbate 80 (available for example as Tween 80™ from Uniqema), can beincluded in an amount of 0% to about 30%, for example about 7% to about30% or about 10% to about 25%, by weight of the carrier. In someembodiments such a surfactant is included in an amount of 0% to about5%, for example 0% to about 2% or 0% to about 1%, by weight of thecarrier.

Conveniently, pre-blended products are available containing a suitablephospholipid+solubilizing agent combination for use in compositions ofthe present invention. Pre-blended phospholipid+solubilizing agentproducts can be advantageous in improving ease of preparation of thepresent compositions.

An illustrative example of a pre-blended phospholipid+solubilizing agentproduct is Phosal 50 PG™, available from Phospholipid GmbH, Germany,which comprises, by weight, not less than 50% phosphatidylcholine, notmore than 6% lysophosphatidylcholine, about 35% propylene glycol, about3% mono- and diglycerides from sunflower oil, about 2% soy fatty acids,about 2% ethanol, and about 0.2% ascorbyl palmitate.

Another illustrative example is Phosal 53 MCT™, also available fromPhospholipid GmbH, which contains, by weight, not less than 53%phosphatidylcholine, not more than 6% lysophosphatidylcholine, about 29%medium chain triglycerides, 3-6% (typically about 5%) ethanol, about 3%mono- and diglycerides from sunflower oil, about 2% oleic acid, andabout 0.2% ascorbyl palmitate (reference composition). A product havingthe above or substantially equivalent composition, whether sold underthe Phosal 53 MCT™ brand or otherwise, is generically referred to hereinas “phosphatidylcholine+medium chain triglycerides 53/29”. A producthaving “substantially equivalent composition” in the present contextmeans having a composition sufficiently similar to the referencecomposition in its ingredient list and relative amounts of ingredientsto exhibit no practical difference in properties with respect toutilization of the product herein.

Yet another illustrative example is Lipoid S75™, available from LipoidGmbH, which contains, by weight, not less than 70% phosphatidylcholinein a solubilizing system. This can be further blended with medium-chaintriglycerides, for example in a 30/70 weight/weight mixture, to providea product (“Lipoid S75™ MCT”) containing, by weight, not less than 20%phosphatidylcholine, 2-4% phosphatidylethanolamine, not more than 1.5%lysophosphatidylcholine, and 67-73% medium-chain triglycerides.

Yet another illustrative example is Phosal 50 SA+™, available fromPhospholipid GmbH, which contains, by weight, not less than 50%phosphatidylcholine and not more than 6% lysophosphatidylcholine in asolubilizing system comprising safflower oil and other ingredients.

The phosphatidylcholine component of each of these pre-blended productsis derived from soy lecithin. Products of substantially equivalentcomposition may be obtainable from other suppliers.

A pre-blended product such as Phosal 50 PG™, Phosal 53 MCT™, Lipoid S75™MCT or Phosal 50 SA+™ can, in some embodiments, constitute substantiallythe entire carrier system (other than the HCA as provided herein). Inother embodiments, additional ingredients are present, for examplemedium-chain mono- and/or diglycerides, ethanol (additional to any thatmay be present in the pre-blended product), a non-phospholipidsurfactant such as polysorbate 80, polyethylene glycol and/or otheringredients. Such additional ingredients, if present, are typicallyincluded in only minor amounts. Illustratively,phosphatidylcholine+medium chain triglycerides 53/29 can be included inthe carrier in an amount of about 50% to 100%, for example about 80% to100%, by weight of the carrier.

Some pre-blended products, including Phosal 50 PG™ and Phosal 53 MCT™,contain a small amount of ascorbyl palmitate, an antioxidant which doesnot meet the definition of an HCA herein. Presence of ascorbyl palmitateor other non-HCA is generally not detrimental, but if desired apre-blended product without such antioxidant can be used as the carrierherein.

In some compositions of the present embodiment, the drug-carrier systemis dispersible in an aqueous phase to form a non-gelling, substantiallynon-transparent liquid dispersion. This property can readily be testedby one of skill in the art, for example by adding 1 part of thedrug-carrier system to about 20 parts of water with agitation at ambienttemperature and assessing gelling behavior and transparency of theresulting dispersion. Compositions having ingredients in relativeamounts as indicated herein will generally be found to pass such a test,i.e., to form a liquid dispersion that does not gel and is substantiallynon-transparent. In “non-gelling” embodiments, the composition does notcontain a gel-promoting agent in a gel-promoting effective amount. Ifgelling behavior is desired, such an agent can be added. A“substantially non-transparent” dispersion is believed to be formed onmixing with an aqueous phase a composition of the invention having anysubstantial amount of the phospholipid component. However, forclarification it is emphasized that compositions of the inventionthemselves, being substantially non-aqueous, are generally clear andtransparent. In this regard, it is noted that phospholipids tend to formbi- and multilamellar aggregates when placed in an aqueous environment,such aggregates generally being large enough to scatter transmittedlight and thereby provide a non-transparent, e.g., cloudy, dispersion.In the case of phosphatidylcholine+medium chain triglycerides 53/29, forexample, dispersion in an aqueous environment typically forms not onlymultilamellar aggregates but also a coarse oil-in-water emulsion.Presence of multilamellar aggregates can often be confirmed bymicroscopic examination in presence of polarized light, such aggregatestending to exhibit birefringence, for example generating acharacteristic “Maltese cross” pattern.

Without being bound by theory, it is believed that behavior of thedrug-carrier system of a composition of the invention upon mixing withan aqueous phase is indicative of how the composition interacts withgastrointestinal fluid following oral administration to a subject.Although formation of a gel can be useful for controlled-release topicaldelivery of a drug, it is believed that gelling would be detrimental toefficient gastrointestinal absorption. For this reason, embodiments ofthe invention described above, wherein the drug-carrier system does notgel when mixed with an aqueous phase, are generally preferred. It isfurther believed, again without being bound by theory, that formation ofbi- and multilamellar aggregates in the gastrointestinal fluid, asevidenced by non-transparency of the dispersion formed upon mixing thedrug-carrier system with an aqueous phase, can be an important factor inproviding the relatively high bioavailability of certain compositions ofthe invention when administered orally.

Carrier ingredients and amounts thereof are selected to providesolubility of the drug in the carrier of at least about 25 mg/ml atabout 25° C.

Illustratively, a drug-carrier system according to the presentembodiment comprises:

about 5% to about 20% by weight ABT-263 free base,

about 15% to about 60% by weight phosphatidylcholine,

about 7% to about 30% by weight medium-chain triglycerides,

about 7% to about 30% by weight medium-chain mono- and diglycerides,

about 7% to about 30% polysorbate 80 surfactant,

about 0.02% to about 0.2% by weight sodium or potassium metabisulfite,

about 0.003% to about 0.01% by weight EDTA or salt thereof, and

about 0.2% to about 0.8% by weight water.

Other excipients can optionally be present in the formulation, so longas they do not adversely affect the storage stability, safety ortherapeutic efficacy of the formulation to an unacceptable degree.However, in a more particular embodiment, the drug-carrier systemconsists essentially of the ingredients listed immediately above.

A prototype formulation of the present embodiment comprises a size 0hard gelatin capsule shell having encapsulated therewithin a liquidsolution that comprises:

about 11% by weight ABT-263 free base,

about 33% by weight phosphatidylcholine,

about 16% by weight medium-chain triglycerides,

about 20% by weight medium-chain mono- and diglycerides,

about 20% by weight polysorbate 80 surfactant,

about 0.05% by weight sodium or potassium metabisulfite,

about 0.005% by weight EDTA or salt thereof, and

about 0.5% by weight water.

The term “about” in descriptions of prototype compositions herein willbe understood to mean that the amounts shown can vary at least withinusual manufacturing tolerances accepted in the pharmaceutical industry.Percentages may not add exactly to 100 because of rounding.

The present invention is not limited by the process used to prepare acomposition as embraced or described herein. Any suitable process ofpharmacy can be used. Illustratively, compositions of the presentembodiment can be prepared by a process comprising simple mixing of therecited ingredients, wherein order of addition is not critical, to forma drug-carrier system. It is noted, however, that if a phospholipidcomponent is used in its solid state, for example in the form of soylecithin, it will generally be desirable to first solubilize thephospholipid with the solubilizing agent component or part thereof.Thereafter other ingredients of the carrier, if any, the drug and theantioxidant can be added by simple mixing, with agitation asappropriate. As mentioned above, use of a pre-blended product comprisingphospholipid and solubilizing agent can simplify preparation of thecomposition. Optionally, the drug-carrier system can be used as a premixfor capsule filling. The term “filling” used in relation to a capsuleherein means placement of a desired amount of a composition in a capsuleshell, and should not be taken to mean that all space in the capsule isnecessarily occupied by the composition.

Where the drug-carrier system comprises a poorly lipid-solublesulfur-containing antioxidant such as sodium or potassium metabisulfite,the process should be adjusted. An illustrative process for preparingsuch a drug-carrier system comprises the following steps.

An API that consists essentially of a compound of Formula I or a saltthereof, for example ABT-263 free base or ABT-263 bis-HCl, is dissolvedin a medium comprising the phospholipid and at least a portion of thesolubilizing agent to provide a lipid solution of the API. As notedabove, a pre-blended product comprising the phospholipid andsolubilizing agent can be used as the medium for dissolution of the API.

Where ABT-263 is to be formulated in its free base form, any solid-stateform of ABT-263 free base can serve as the API. However, it willgenerally be found preferable to use a crystalline form of ABT-263 freebase as API, for example a solvated or non-solvated crystalline form. Ina particular embodiment of the present process, a non-solvatedcrystalline form such as Form I or Form II crystalline ABT-263 asdescribed herein is used as API.

A non-phospholipid surfactant and, optionally, the balance of thesolubilizing agent, is admixed with the solubilizing agent (prior to orsimultaneously with dissolution of the API) or with the lipid solution(after dissolution of the API). As noted above, the non-phospholipidsurfactant is illustratively a polysorbate such as polysorbate 80. Thebalance of the solubilizing agent can be the same material as theportion of solubilizing agent used together with the phospholipid todissolve the API; alternatively it can be a different material. Forexample, the portion of solubilizing agent used together with thephospholipid for dissolution of the API can comprise one or moremedium-chain triglycerides, and the balance of solubilizing agentadmixed in the present step can comprise one or more medium-chain mono-and/or diglycerides, for example a caprylic/capric mono- and diglycerideproduct such as Imwitor 742™.

Separately, a poorly lipid-soluble sulfur-containing antioxidant isdissolved in water to prepare an aqueous stock solution. Stock solutionsat about 10% to about 18% by weight concentration will generally befound suitable, as explained above.

The aqueous stock solution is then admixed with the lipid solution,typically after addition of the non-phospholipid surfactant, to providea liquid solution for encapsulation.

Optionally, the resulting liquid solution is encapsulated in a capsuleshell by any known encapsulation process.

Second Composition Embodiment

A composition of the second embodiment set forth hereinabove comprises acapsule shell having encapsulated therewithin, in an amount not greaterthan about 1000 mg per capsule, a liquid solution of a compound ofFormula I or a pharmaceutically acceptable salt thereof in a free baseequivalent amount of at least about 2.5% by weight of the solution, in asubstantially non-ethanolic carrier that comprises as pharmaceuticallyacceptable excipients:

-   -   (a) at least one phospholipid,    -   (b) at least one solubilizing agent for the at least one        phospholipid, selected from the group consisting of glycols,        glycolides, glycerides and mixtures thereof,    -   (c) at least one non-phospholipid surfactant, and    -   (d) a pharmaceutically acceptable HCA.

In a capsule of the present embodiment, ABT-263 is “in solution” in theencapsulated liquid as in a composition of the first embodimentdescribed above. The encapsulated liquid is “substantiallynon-ethanolic”, i.e., having no ethanol, or having an amount of ethanolthat is small enough to be, in practical terms, essentiallynon-deleterious to performance or properties of the capsule. Moreparticularly, any ethanol that is present must be below a thresholdconcentration at which integrity of the capsule shell is compromised.Typically the encapsulated liquid comprises zero to less than about 5%by weight ethanol. This is especially important where a hard capsuleshell, for example a hard gelatin or hydroxypropylmethylcellulose (HPMC)capsule shell, is used. Soft capsule shells, for example soft gelatin orstarch-based shells containing a plasticizer, can tolerate somewhathigher amounts of ethanol. Certain pre-blended phospholipid productsuseful herein contain small amounts of ethanol that are non-deleteriouseven to a hard gelatin capsule; for example Phosal 53 MCT™ can containup to about 6% ethanol. When used illustratively in an amount notexceeding about 75% by weight of the encapsulated liquid, Phosal 53 MCT™is seen to contribute ethanol in an amount not exceeding about 4.5% byweight of the encapsulated liquid, which remains “substantiallynon-ethanolic” as defined herein.

In most embodiments, the encapsulated liquid is also “substantiallynon-aqueous”, as defined above in relation to compositions of the firstembodiment.

As indicated above, the encapsulated liquid comprises, inter alia, aphospholipid, and a pharmaceutically acceptable solubilizing agent forthe phospholipid. The solubilizing agent, or the combination ofsolubilizing agent and phospholipid, may also assist in solubilizing theABT-263, as may other ingredients, such as a non-phospholipidsurfactant. Phospholipids and solubilizing agents, including pre-blendedproducts, useful herein are as described above in relation tocompositions of the first embodiment.

Illustratively, a suitable amount of phospholipid in the encapsulatedliquid of the present embodiment is about 15% to about 60%, for exampleabout 20% to about 45%, by weight of the encapsulated liquid, althoughgreater and lesser amounts can be useful in particular situations.

If the solubilizing agent comprises one or more glycols, these canillustratively present in a total glycol amount of at least about 1% butless than about 50%, for example less than about 30%, less than about20%, less than about 15% or less than about 10% by weight of thecarrier. In some embodiments, the carrier comprises substantially noglycol.

Where one or more glycerides are present as a major component of thesolubilizing agent, a suitable total amount of glycerides is an amounteffective to solubilize the phospholipid and, in combination with otherexcipients, effective to maintain the compound of Formula I or saltthereof, for example ABT-263 free base or ABT-263 bis-HCl, in solution.For example, glycerides such as medium-chain mono-, di- andtriglycerides can be present in a total glyceride amount of about 15% toabout 60%, for example about 20% to about 45%, by weight of theencapsulated liquid, although greater and lesser amounts can be usefulin particular situations. In one embodiment, the encapsulated liquidcomprises about 7% to about 30%, for example about 10% to about 25%, byweight medium-chain triglycerides and about 7% to about 30%, for exampleabout 10% to about 25%, by weight medium-chain mono- and diglycerides.

The encapsulated liquid of the present embodiment further comprises apharmaceutically acceptable non-phospholipid surfactant, for example asdescribed above in relation to compositions of the first embodiment.Illustratively, a surfactant such as a polysorbate, e.g., polysorbate80, can be included in an amount of about 7% to about 30%, for exampleabout 10% to about 25%, by weight of the encapsulated liquid.

Illustratively, the encapsulated liquid solution according to thepresent embodiment comprises:

about 5% to about 20% by weight ABT-263 free base,

about 15% to about 60% by weight phosphatidylcholine,

about 7% to about 30% by weight medium-chain triglycerides,

about 7% to about 30% by weight medium-chain mono- and diglycerides,

about 7% to about 30% polysorbate 80 surfactant,

about 0.02% to about 0.2% by weight sodium or potassium metabisulfite,

about 0.003% to about 0.01% by weight EDTA or salt thereof, and

about 0.2% to about 0.8% by weight water.

Other excipients can optionally be present in the encapsulated solution,so long as they do not adversely affect the storage stability, safety ortherapeutic efficacy of the capsule to an unacceptable degree. However,in a more particular embodiment, the encapsulated liquid solutionconsists essentially of the ingredients listed immediately above.

The capsule shell can be of any pharmaceutically acceptable material,including hard or soft gelatin. A capsule shell size is selectedappropriate to the amount of liquid to be encapsulated. For example, asize 0 capsule shell can be used to encapsulate up to about 600 mg ofliquid and a size 00 capsule shell up to about 900 mg of liquid.

A prototype capsule of the present invention comprises a size 0 hardgelatin capsule shell having encapsulated therewithin a liquid solutionthat comprises:

about 50 mg ABT-263 free base,

about 150 mg phosphatidylcholine,

about 75 mg medium-chain triglycerides,

about 90 mg medium-chain mono- and diglycerides,

about 90 mg polysorbate 80 surfactant,

about 0.25 mg sodium or potassium metabisulfite,

about 0.025 mg EDTA or salt thereof, and

about 2.5 mg water.

Illustratively, a capsule of the invention can be prepared by a processcomprising simple mixing of the recited ingredients, wherein order ofaddition is not critical, to form a liquid solution for encapsulation,followed by encapsulation of the liquid in a hard or soft gelatincapsule shell to form a capsule. It is noted, however, that if thephospholipid is used in its solid state, for example in the form of soylecithin, it will generally be desirable to first solubilize thephospholipid with the solubilizing agent or part thereof. Thereafterother excipients and the ABT-263 can be added by simple mixing, withagitation as appropriate. Use of a pre-blended product comprisingphospholipid and solubilizing agent can simplify preparation of thecomposition. For example, the phospholipid can comprisephosphatidylcholine and the solubilizing agent pre-blended therewith cancomprise medium-chain triglycerides, as in the case of Phosal 53 MCT™ orLipoid S75™ MCT. Illustratively, the pre-blended product comprises about50% to about 75% phosphatidylcholine and about 15% to about 30%medium-chain triglycerides.

Where the solution for encapsulation comprises a poorly lipid-solublesulfur-containing antioxidant such as sodium or potassium metabisulfite,the process should be adjusted. An illustrative process for preparingsuch a solution is as described above in relation to a composition ofthe first embodiment. The resulting liquid solution is then encapsulatedin a capsule shell by any known encapsulation process.

Third Composition Embodiment

A composition of the third embodiment set forth hereinabove comprises anorally deliverable liquid pharmaceutical composition comprising anaqueous medium having suspended therein a solid particulate compoundhaving a D₉₀ particle size not greater than about 3 μm; wherein thecompound is of Formula I or a pharmaceutically acceptable salt thereof,for example ABT-263 free base or ABT-263 bis-HCl, and is present in afree base equivalent amount of at least about 2.5% by weight of thecomposition; and wherein the aqueous medium further comprises at leastone pharmaceutically acceptable surfactant and at least onepharmaceutically acceptable basifying agent in amounts that areeffective together to inhibit particle size increase.

A suspension composition in accordance with the present embodimentcomprises a nanosized solid particulate drug compound. It is found thatin the suspensions described herein the drug nanoparticles do notappreciably agglomerate, resulting in production of stable formulations.

Unless the context demands otherwise, the term “nanoparticle” as usedherein means a particle of size (i.e., diameter in the longest dimensionof the particle) not greater than about 3 μm (3,000 nm). “Nanoparticles”as recited herein therefore include not only “submicron” particles,i.e., having a size less than about 1 μm, but also “micron-sized”particles of about 1 to about 3 μm. Likewise, the adjective “nanosized”as used herein refers to nanoparticles as defined immediately above.Unless the context demands otherwise, the term “nanoparticulate” asapplied to a suspension or other composition herein, and likewise theterm “nanosuspension”, means having a D₉₀ particle size not greater thanabout 3 μm.

The D₉₀ particle size of a composition is a parameter such that 90% byvolume of particles in the composition are smaller in their longestdimension than that parameter, as measured by any conventional particlesize measuring technique known to those skilled in the art. Suchtechniques include, for example, sedimentation field flow fractionation,photon correlation spectroscopy, light scattering, and diskcentrifugation. In various compositions of the present embodiment,suspensions are provided having a D₉₀ particle size not greater thanabout 3,000 nm, not greater than about 2,000 nm, not greater than about1,500 nm, not greater than about 1,000 nm, not greater than about 900nm, not greater than about 800 nm, not greater than about 700 nm, notgreater than about 600 nm or not greater than about 500 nm.

The D₅₀ particle size of a composition is a parameter such that 50% byvolume of particles in the composition are smaller in their longestdimension than that parameter, as measured by any conventional particlesize measuring technique known to those skilled in the art. D₅₀ particlesize is therefore a measure of volume median particle size but issometimes referred to as “average” or “mean” particle size. In variouscompositions of the present embodiment, suspensions are provided havinga D₅₀ particle size not greater than about 1,000 nm, not greater thanabout 900 nm, not greater than about 800 nm, not greater than about 700nm, not greater than about 600 nm, not greater than about 500 nm, notgreater than about 400 nm, not greater than about 350 nm or not greaterthan about 300 nm.

In some cases, a suspension as provided herein has a D₉₀ particle sizenot greater than about 1,000 nm and a D₅₀ particle size not greater thanabout 400 nm. In other cases, a suspension as provided herein has a D₉₀particle size not greater than about 800 nm and a D₅₀ particle size notgreater than about 350 nm.

The terms “low solubility” and “poorly soluble” as used in relation tocompositions of the present embodiment refer to a solubility in waternot greater than about 100 μg/ml. The present invention can beespecially advantageous for drugs that are essentially insoluble inwater, i.e., having a solubility of less than about 10 μg/ml. It isbelieved, without being bound by theory, that the advantages ofnanoparticulate suspensions for such drugs arise in part not only fromimproved dissolution rate, which is proportional to surface areaaccording to the well known Whitney-Noyes equation, but also fromimproved solubility according to the Kelvin equation. This can result inenhanced bioavailability as well as potentially reduce food effect.

The nanoparticulate suspension comprises a compound of Formula I or asalt thereof as a discrete solid-state phase that can be crystalline,semi-crystalline or amorphous. In the case of ABT-263, the free baseform of which, as prepared according to the '135 publication, is anamorphous or glassy solid, it is generally preferred to use acrystalline salt form of the drug, such as for example ABT-263 bis-HCl,in preparing the nanosuspension. However, upon suspension of the salt inpresence of a basifying agent such as sodium bicarbonate, someconversion of salt to free base can occur, resulting in the solid-statephase becoming at least partly amorphous. Accordingly, in oneembodiment, the nanosuspension comprises ABT-263 free base, ABT-263bis-HCl or a combination thereof. Despite the likelihood that the drugparticles in an ABT-263 nanosuspension are at least partly amorphous, aremarkably high degree of physical stability has been observed in such ananosuspension, as illustrated in Example 14 below.

It has been found that nanoparticulate suspensions as described hereinoffer not only the advantage of physical stability providing acceptableproduct shelf life, but also the robustness of manufacturing processthat is desirable for a commercial product.

The concentration of drug in the suspension is at least about 25 mg/ml,e.g., about 25 to about 500 mg/ml. Illustratively, for example where thedrug is ABT-263, the drug concentration in various embodiments is about25 to about 400 mg/ml, for example about 25, about 30, about 40, about50, about 75, about 100, about 125, about 150 or about 200 mg/ml, byfree base equivalent weight.

Compositions of the present invention have good storage-stabilityproperties. In particular, they are physically stable, at least in thatthey do not have an unacceptable tendency to undergo particle sizeincrease over time, for example through particle agglomeration. Particleagglomeration is a common problem in nanoparticulate suspensions.Surface modifying agents such as surfactants are important in reducingthe tendency of nanoparticles to agglomerate; the at least onesurfactant present in a composition of the present invention isbelieved, without being bound by theory, to help in this regard.

A “basifying agent” herein is any agent that raises the pH of thesuspension medium. Any pharmaceutically acceptable basifying agent canbe used, including without limitation hydroxides and bicarbonates ofalkali metals such as sodium and potassium. The invention is illustratedherein with particular reference to sodium bicarbonate, but it will berecognized that other basifying agents can be substituted for sodiumbicarbonate if desired.

Amount of sodium bicarbonate useful in a composition of the invention isnot narrowly critical, and one of ordinary skill in the art can readilyoptimize the amount for any particular composition, for example byroutine storage-stability testing. In general, good results can beobtained with sodium bicarbonate in an amount of about 20 to about 200mg/ml, for example about 40 to about 160 mg/ml.

The choice and amount of surfactant is likewise not narrowly critical,and is likely to depend to some extent on the particular drug compoundto be formulated and the drug loading desired. Non-limiting examples ofsurfactants include, either individually or in combination, quaternaryammonium compounds, for example benzalkonium chloride, benzethoniumchloride and cetylpyridinium chloride; dioctyl sodium sulfosuccinate;polyoxyethylene alkylphenyl ethers, for example nonoxynol 9, nonoxynol10 and octoxynol 9; poloxamers (polyoxyethylene and polyoxypropyleneblock copolymers), for example poloxamer 188 and poloxamer 237;polyoxyethylene fatty acid glycerides and oils, for examplepolyoxyethylene (8) caprylic/capric mono- and diglycerides,polyoxyethylene (35) castor oil and polyoxyethylene (40) hydrogenatedcastor oil; polyoxyethylene alkyl ethers, for example ceteth-10,laureth-4, laureth-23, oleth-2, oleth-10, oleth-20, steareth-2,steareth-10, steareth-20, steareth-100 and polyoxyethylene (20)cetostearyl ether; polyoxyethylene fatty acid esters, for examplepolyoxyethylene (20) stearate, polyoxyethylene (40) stearate andpolyoxyethylene (100) stearate; sorbitan esters, for example sorbitanmonolaurate, sorbitan monooleate, sorbitan monopalmitate and sorbitanmonostearate; polyoxyethylene sorbitan esters, for example polysorbate20 and polysorbate 80; propylene glycol fatty acid esters, for examplepropylene glycol laurate; sodium lauryl sulfate; fatty acids and saltsthereof, for example oleic acid, sodium oleate and triethanolamineoleate; glyceryl fatty acid esters, for example glyceryl monooleate,glyceryl monostearate and glyceryl palmitostearate; α-tocopherylpolyethylene glycol succinate (TPGS); tyloxapol; and the like. In oneembodiment, the at least one surfactant is a poloxamer or mixture ofpoloxamers. Poloxamer 188 is a specific example. One or more surfactantstypically constitute in total about 10 to about 100 mg/ml of thesuspension. In the case of poloxamer 188, an illustratively suitableamount is about 10 to about 100 mg/ml, for example about 15 to about 60mg/ml, of the suspension.

The aqueous medium of the suspension can take the form of water, anaqueous injectable fluid such as saline (e.g., phosphate-buffered salineor PBS) or an imbibable liquid such as fruit juice or a carbonatedbeverage. In one embodiment the nanoparticulate drug compound, the atleast one surfactant and at least one basifying agent (and optionallyadditional ingredients) are prepared as a dry powder mix forreconstitution with a suitable aqueous medium to form a suspensioncomposition of the invention shortly before use. Such a reconstitutablepowder should contain, in addition to the ingredients recited above, atleast one pharmaceutically acceptable dispersant or bulking agent,typically a water-soluble material such as a sugar, e.g., dextrose,mannitol or dextran; a phosphate salt, e.g., sodium or potassiumphosphate; an organic acid, e.g., citric acid or tartaric acid, or asalt thereof; or a mixture of such materials. A dry powder mix canalternatively be administered to a subject for resuspension of thenanoparticles in the gastrointestinal fluid; for such administration thepowder mix can if desired be formed into a tablet or filled into acapsule.

It is desirable to provide a formulation that is not only physicallystable but also chemically stable. More particularly, such a formulationshould not exhibit an unacceptable degree of oxidative degradation ofthe compound of Formula I, for example at the thioether linkage of the(phenylsulfanyl)methyl group thereof.

In this regard, a composition of the present invention containing acompound of Formula I such as ABT-263 free base, ABT-263 bis-HCl or acombination thereof possesses a significant advantage over solutioncompositions of ABT-263 previously disclosed in the art, for example inthe '135 publication or in Tse et al. (2008), supra. The solid-stateform (whether crystalline, semi-crystalline or amorphous) of ABT-263present in a nanosuspension as provided herein is believed to besignificantly more resistant to oxidative degradation than ABT-263 insolution.

However, if desired, any remaining tendency for oxidative degradationcan be further reduced by inclusion of a suitable antioxidant, moreparticularly an HCA as described hereinabove in the suspensioncomposition.

In view of the aqueous nature of the suspension medium, water-solubleinorganic antioxidants of the sulfite, bisulfite, metabisulfite andthiosulfate classes can be particularly useful. Such antioxidants can beincluded in any suitable amount, for example about 0.02% to about 2%, orabout 0.05% to about 1%, by weight, of the composition.

Sodium and potassium salts of sulfites, bisulfites, metabisulfites andthiosulfates are especially useful antioxidants according to the presentembodiment; more particularly sodium and potassium metabisulfites.

To further minimize sulfoxide formation, a chelating agent such as EDTAor a salt thereof (e.g., disodium EDTA or calcium disodium EDTA) isoptionally added, for example in an amount of about 0.002% to about 0.2%by weight of the composition.

Other optional ingredients of the suspension composition includebuffers, coloring agents, flavoring agents, preservatives, sweeteners,tonicifying agents and combinations thereof.

A process for preparing a nanoparticulate pharmaceutical composition ofthe present embodiment comprises providing an API that comprises acompound of Formula I or a pharmaceutically acceptable salt thereof, forexample ABT-263 or a crystalline salt thereof; wet-milling the API inpresence of at least one basifying agent, such as sodium bicarbonate, toa D₉₀ particle size not greater than about 3 μm to provide a milled drugsubstance; and suspending the milled drug substance in an aqueous mediumwith the aid of at least one surfactant; wherein the at least onebasifying agent and the at least one surfactant are present in theresulting suspension in amounts that are effective together to inhibitparticle size increase.

Any suitable wet-milling process can be used. A particular wet-millingprocess that has been found useful is high-pressure homogenization asillustratively described in Example 13 below.

The present invention is not limited to compositions prepared by anyprocess described herein; however, a composition prepared by the aboveprocess is a particular embodiment of the invention.

In one embodiment, the process further comprises adding at least onepharmaceutically acceptable dispersant or bulking agent to thesuspension, drying (for example freeze-drying or lyophilizing, oralternatively spray-drying) the suspension to provide a reconstitutabledry powder, and optionally forming the powder into a tablet (for exampleby molding or compression) or filling the powder into a capsule, toprepare a unit dosage form.

In addition to the stabilizing benefits of sodium bicarbonate, it isfound that in presence of sodium bicarbonate, wet-milling to smallerparticle sizes, for example to a D₉₀ particle size not greater thanabout 700 nm, is possible. Without sodium bicarbonate, as illustrativelyshown in Example 14 hereinbelow, using the same processing parameters,D₉₀ particle size can not be reduced below about 1,000 nm. Thewet-milling method used in the present process has the advantage, bycomparison with dry-milling, that it reduces exposure of the API to hightemperature and thereby reduces risk of thermal decomposition of theAPI. In one embodiment, processing temperature is controlled, forexample within about 1 to about 5 degrees of a target temperature ofabout 5° C. to about 30° C. This can be achieved by conventional means,such as by running the formulation through a heat exchanger immersed ina chilled water bath.

The composition can be prepared for wet-milling at its finalconcentration, or it can be prepared at higher concentration and dilutedto a desired concentration after wet-milling. The at least onesurfactant and, if desired, optional additional ingredients, can beadded before or after wet-milling.

Fourth Composition Embodiment

A composition of the fourth embodiment set forth hereinabove comprisesan orally deliverable solid dispersion comprising, in essentiallynon-crystalline, for example amorphous, form, a compound of Formula I ora pharmaceutically acceptable salt thereof in a free base equivalentamount of at least about 2.5% by weight of the composition, dispersed ina solid matrix that comprises (a) a pharmaceutically acceptablewater-soluble polymeric carrier and (b) a pharmaceutically acceptablesurfactant.

A solid dispersion in accordance with the present embodiment comprises acompound of Formula I or a pharmaceutically acceptable salt thereof, forexample ABT-263 free base or ABT-263 bis-HCl, in an essentiallynon-crystalline or amorphous form, which is usually more soluble thanthe crystalline form. The term “solid dispersion” herein encompassessystems having small solid-state particles of one phase dispersed inanother solid-state phase. More particularly, the present soliddispersions comprise one or more active ingredients dispersed in aninert carrier or matrix in solid state, and can be prepared by meltingor solvent methods or by a combination of melting and solvent methods.According to the present embodiment a solvent method as described hereinis particularly favored, avoiding the risk of thermal decomposition ofthe active ingredient by exposure to temperatures required to melt thepolymeric carrier.

An “amorphous form” refers to a particle without definite structure,i.e., lacking crystalline structure.

The term “essentially non-crystalline” herein means that no more thanabout 5%, for example no more than about 2% or no more than about 1%crystallinity is observed by X-ray diffraction analysis. In a particularembodiment, no detectable crystallinity is observed by one or both ofX-ray diffraction analysis or polarization microscopy.

ABT-263 bis-HCl, by virtue of its crystalline nature, is typically moreconvenient to use as an API than ABT-263 free base, which as preparedaccording to the '135 publication is an amorphous or glassy solid.However, there may be advantages in providing a solid dispersionformulation of ABT-263 wherein the ABT-263 is in free base form, as thedrug will be less susceptible to crystallization within the formulationor immediately upon release therefrom. Thus in a particular embodiment,the composition comprises ABT-263 free base. It is emphasized that, inthis embodiment, it is not necessarily the free base form of ABT-263that is used as the API in preparing the composition.

The concentration of drug in the solid dispersion of the presentembodiment is at least about 2.5%, e.g., about 2.5% to about 50%, byfree base equivalent weight. Illustratively, for example where the drugis ABT-263, the drug concentration in various compositions is at leastabout 5%, e.g., about 5% to about 40%, for example about 5%, about 10%,about 15%, about 20%, about 25%, about 30%, about 35% or about 40%, byfree base equivalent weight.

The major component of the matrix of a solid dispersion product is apolymer that is hydrophilic or water-soluble at least in a part of thepH scale, more particularly at a pH occurring in the gastrointestinal(GI) tract, or a combination of such polymers. A polymer or polymermixture useful herein is solid at ambient temperature and, in theinterests of good storage stability at a range of temperatures, shouldremain solid even at the highest temperatures typically experiencedduring storage, transport and handling of the product. A useful propertyof a polymer determining its usefulness herein is therefore its glasstransition temperature (T_(g)). Suitable water-soluble polymers include,but are not limited to, those having a T_(g) of at least about 50° C.,more particularly about 80° C. to about 180° C. Methods for determiningT_(g) values of organic polymers are described for example in Sperling,ed. (1992) Introduction To Physical Polymer Science, 2nd edition, JohnWiley & Sons, Inc.

Non-limiting examples of polymeric carriers useful herein include:

-   -   homopolymers and copolymers of N-vinyl lactams, especially        homopolymers and copolymers of N-vinyl pyrrolidone, e.g., the        homopolymer polyvinylpyrrolidone (PVP or povidone) and        copolymers such as those comprising monomers of N-vinyl        pyrrolidone and vinyl acetate (copovidone) or N-vinyl        pyrrolidone and vinyl propionate;    -   cellulose esters and cellulose ethers, in particular        methylcellulose, ethylcellulose, (hydroxyalkyl)celluloses such        as hydroxypropylcellulose, (hydroxyalkyl)alkyl-celluloses such        as hydroxypropylmethylcellulose (HPMC or hypromellose),        cellulose phthalates and succinates such as cellulose acetate        phthalate, hydroxypropylmethylcellulose phthalate,        hydroxypropylmethylcellulose succinate and        hydroxypropylmethylcellulose acetate succinate (HPMC-AS);    -   high molecular weight polyalkylene oxides such as polyethylene        oxide, polypropylene oxide and copolymers of ethylene oxide and        propylene oxide (poloxamers);    -   polyacrylates and polymethacrylates such as methacrylic        acid/ethyl acrylate copolymers, methacrylic acid/methyl        methacrylate copolymers, butyl methacrylate/2-dimethylaminoethyl        methacrylate copolymers, poly(hydroxyalkyl acrylates) and        poly(hydroxyalkyl methacrylates);    -   polyacrylamides;    -   vinyl acetate polymers such as copolymers of vinyl acetate and        crotonic acid, partially hydrolyzed polyvinyl acetate (also        referred to as partially saponified “polyvinyl alcohol”) and        polyvinyl alcohol;    -   oligo- and polysaccharides such as carrageenans, galactomannans        and xanthan gum;        and mixtures of two or more thereof.

In some compositions, the solid dispersion matrix comprises one or morepolymeric carriers selected from the group consisting of copovidone,povidone and HPMC-AS. A particular example of a useful copovidone is oneconsisting of about 60% N-vinyl pyrrolidone and about 40% vinyl acetatemonomers. A particular example of a useful povidone is one having aK-value (a measure of viscosity of an aqueous solution of the povidone)of about 30.

One or more polymeric carriers typically constitute in total about 20%to about 90%, for example about 40% to about 85%, by weight of the soliddispersion.

Upon oral administration and exposure to GI fluid, it is believedwithout being bound by theory that, through interplay between thepolymeric carrier and a surfactant component of the solid dispersion, asuitable release rate and inhibition of crystallization orrecrystallization of the active ingredient are provided, therebypermitting bioabsorption.

Particularly useful as surfactants in solid dispersions of the presentembodiment are pharmaceutically acceptable non-ionic surfactants,especially those having a hydrophilic-lipophilic balance (HLB) value ofabout 12 to about 18, for example about 13 to about 17, or about 14 toabout 16. The HLB system (see Fiedler (2002) Encyclopedia of Excipients,5th edition, Aulendorf: ECV-Editio-Cantor-Verlag) attributes numericvalues to surfactants, with lipophilic substances receiving lower HLBvalues and hydrophilic substances receiving higher HLB values.

Non-limiting examples of non-ionic surfactants useful in compositions ofthe present embodiment include:

-   -   polyoxyethylene castor oil derivatives such as PEG-35 castor oil        (e.g., Cremophor EL™ of BASF Corp. or equivalent product),        PEG-40 hydrogenated castor oil (e.g., Cremophor RH 40™ or        equivalent product) and PEG-60 hydrogenated castor oil (e.g.,        Cremophor RH™ 60 or equivalent product);    -   fatty acid monoesters of sorbitan, for example sorbitan        monooleate (e.g., Span™ 80 or equivalent product), sorbitan        monostearate (e.g., Span™ 60 or equivalent product), sorbitan        monopalmitate (e.g., Span™ 40 or equivalent product) and        sorbitan monolaurate (e.g., Span™ 20 or equivalent product);    -   fatty acid monoesters of polyoxyethylene sorbitan (polysorbates)        such as PEG-20 sorbitan monooleate (polysorbate 80, e.g., Tween™        80 or equivalent product) PEG-20 sorbitan monostearate        (polysorbate 60, e.g., Tween™ 60 or equivalent product), PEG-20        sorbitan monopalmitate (polysorbate 40, e.g., Tween™ 40 or        equivalent product), or PEG-20 sorbitan monolaurate (polysorbate        20, e.g., Tween™ 20 or equivalent product);    -   poloxamers such as poloxamer 124, poloxamer 188, poloxamer 237,        poloxamer 388 or poloxamer 407;    -   α-tocopheryl polyethylene glycol succinate (TPGS or vitamin E        polyethylene glycol succinate, see U.S. National Formulary);        and mixtures of two or more thereof.

One or more surfactants typically constitute in total about 2% to about25%, for example about 5% to about 20%, by weight of the soliddispersion.

A dosage form of the present embodiment can consist of, or consistessentially of, a solid dispersion as described above. However, in somecases a dosage form of the present embodiment contains additionalexcipients and requires additional processing of the solid dispersion.For example, the solid dispersion can be ground to a powder and filledinto a capsule shell or molded or compressed to form a tablet, withadditional excipients as may be conventionally used in such dosageforms.

Thus orally deliverable solid dosage forms of the present embodimentinclude but are not limited to capsules, dragees, granules, pills,powders and tablets. Excipients commonly used to formulate such dosageforms include encapsulating materials or formulation additives such asabsorption accelerators, antioxidants, binders, buffers, coating agents,coloring agents, diluents, disintegrating agents, emulsifiers,extenders, fillers, flavoring agents, humectants, lubricants,preservatives, propellants, releasing agents, sterilizing agents,sweeteners, solubilizers and mixtures thereof. Examples of specificexcipients include agar, alginic acid, aluminum hydroxide, benzylbenzoate, 1,3-butylene glycol, castor oil, cellulose, cellulose acetate,cocoa butter, corn starch, corn oil, cottonseed oil, ethanol, ethylacetate, ethyl carbonate, ethyl cellulose, ethyl laureate, ethyl oleate,gelatin, germ oil, glucose, glycerol, groundnut oil, isopropanol,isotonic saline, lactose, magnesium hydroxide, magnesium stearate, malt,olive oil, peanut oil, potassium phosphate salts, potato starch,propylene glycol, talc, tragacanth, water, safflower oil, sesame oil,sodium carboxymethyl cellulose, sodium lauryl sulfate, sodium phosphatesalts, soybean oil, sucrose, tetrahydrofurfuryl alcohol, and mixturesthereof.

A solvent process for preparing a solid dispersion as described abovecomprises dissolving the API, the polymeric carrier and the surfactantin a suitable solvent; and removing the solvent to provide the soliddispersion. Optionally, where the API is in salt form and it is desiredto provide a solid dispersion of the drug in free base form, a base isadded before solvent removal to effect conversion of the API to itscorresponding free base. For example, where the API is ABT-263 bis-HCl,addition of a base such as sodium hydroxide (NaOH), potassium hydroxide(KOH), sodium bicarbonate (NaHCO₃), potassium bicarbonate (KHCO₃) orammonium bicarbonate (NH₄HCO₃) in an amount of at least 2 moles per moleof API can result in conversion of the API to ABT-263 free base. Theinorganic salt by-product, illustratively NaCl, KCl or NH₄Cl, can remainin the product or is optionally extracted before solvent removal.

In the dissolving step, the various components can be added in anyorder. For example, each ingredient can be added to the solventseparately and then dissolved therein. Alternatively, the polymericcarrier and/or surfactant can be pre-mixed with the API, and theresulting mixture then added to the solvent. However, it will generallybe found convenient, when the process includes in situ salt-to-free baseconversion, to first add the API salt and the base to the solvent, then(optionally after extraction of a salt by-product) add the polymericcarrier and surfactant.

In principle any solvent can be used so long as it is effective todissolve the active ingredient, polymer carrier and surfactant.Non-limiting examples of solvents that can be useful include methanol,ethanol, acetone and mixtures thereof. Optionally a cosolvent can beincluded.

Where it is desired to extract a salt by-product such as NaCl, KCl orNH₄Cl prior to solvent removal, a solvent can be selected wherein thesalt by-product is insoluble, thereby permitting extraction of the saltby-product by filtration.

Solvent removal can be accomplished using heat, vacuum or a combinationthereof. If heat is used, it is generally preferable to avoid exceedingthe glass transition temperature (T_(g)) of the polymeric matrix. Formost purposes heating at a temperature of about 50° C. to about 80° C.,for example about 55° C. to about 75° C., will be found suitable. Aftersolvent removal, the resulting product is cooled (if necessary) toambient temperature.

Further process details can be found in the illustrative processes ofExamples 16 and 17 below.

Fifth Composition Embodiment

A composition of the fifth embodiment set forth hereinabove comprises anorally deliverable pharmaceutical dosage form comprising a soliddispersion or solid solution that comprises (a) a compound of Formula Ior a pharmaceutically acceptable salt thereof in a free base equivalentamount of at least about 2.5% by weight of the composition, (b) at leastone pharmaceutically acceptable polymer and (c) at least onepharmaceutically acceptable solubilizer.

In dosage forms of the present embodiment, the active ingredient ispresent as a solid dispersion or as a solid solution. The term “soliddispersion” in relation to the present embodiment defines a system in asolid state (as opposed to a liquid or gaseous state) comprising atleast two components, wherein one component is dispersed evenlythroughout the other component or components. For example, the activeingredient or combination of active ingredients is dispersed in a matrixcomprising the pharmaceutically acceptable polymer(s) andpharmaceutically acceptable solubilizers. The term “solid dispersion”encompasses systems having small particles, typically less than 1 μm indiameter, of one phase dispersed in another phase. When said dispersionis such that the system is chemically and physically uniform orhomogeneous throughout or consists of one phase (as defined inthermodynamics), such a solid dispersion will be called a “solidsolution” or a “glassy solution”. A glassy solution is a homogeneous,glassy system in which a solute is dissolved in a glassy solvent. Glassysolutions and solid solutions are preferred physical systems accordingto the present embodiment. These systems do not contain any significantamount of active ingredients in a crystalline or microcrystalline state,as evidenced by thermal analysis (DSC) or X-ray diffraction analysis(WAXS).

Dosage forms according to the present embodiment are characterized byexcellent stability and, in particular, exhibit high resistance againstrecrystallization or decomposition of the active ingredient(s).

Dosage forms of the present embodiment exhibit a release and absorptionbehavior that is characterized by relatively high attainable AUC,relatively high attainable C_(max), and relatively low T_(max).

A dispersion formed upon contact of a dosage form of the presentembodiment with an aqueous liquid may also be useful as such, forexample as an oral liquid dosage form or a parenteral injection.

Generally, the solid dispersion product of the present embodimentcomprises

-   -   (a) about 2.5% to about 40%, preferably about 2.5% to about 25%,        by weight of a compound of Formula I or a salt thereof, for        example ABT-263 free base, ABT-263 bis-HCl or ABT-263 sodium        salt,    -   (b) about 40% to about 95%, preferably about 50% to about 94%,        by weight of at least one pharmaceutically acceptable polymer,    -   (c) about 2% to about 20%, preferably about 5% to about 20%, by        weight of at least one solubilizer, and    -   (d) zero to about 15%, preferably zero to 10%, by weight of        additives.

Whereas the dosage form of the present embodiment may consist entirelyof solid dispersion product, additives and adjuvants can be used informulating the solid dispersion product into the dosage form.Generally, the dosage form comprises at least about 10%, preferably atleast about 40%, and most preferably at least about 45%, by weight ofsolid dispersion product, based on the total weight of the solid dosageform.

Typically, a single dosage form of the present embodiment contains about50 mg to about 1000 mg, preferably about 75 mg to about 600 mg, inparticular about 100 mg to about 500 mg, of free base equivalent of acompound of Formula I, for example ABT-263, or a salt thereof.

In suitable embodiments, the active ingredient is selected from thegroup consisting of the free base, the sodium salt and thebis-hydrochloride salt of ABT-263, and combinations thereof. In apreferred embodiment the active ingredient is ABT-263 free base.

The term “solubilizer” as used in relation to the present embodimentrefers to a pharmaceutically acceptable nonionic or anionic surfactant.The solubilizer may effect an instantaneous emulsification of the activeingredient released from the dosage form and/or prevent precipitation ofthe active ingredient in the aqueous fluid of the gastrointestinaltract. A single solubilizer or combination of solubilizers may be used.The solubilizer may be selected from the group consisting of nonionicsolubilizers, anionic solubilizers and combinations thereof. In somecompositions of the present embodiment, the solid dispersion productcomprises a combination of two or more pharmaceutically acceptablesolubilizers.

Illustratively, a nonionic solubilizer can be selected from the groupconsisting of polyol fatty acid esters, polyalkoxylated polyol fattyacid esters, polyalkoxylated fatty alcohol ethers, tocopheryl compoundsor mixtures of two or more thereof, and an anionic solubilizer can beselected from the group consisting of alkyl sulfates, alkylcarboxylates,alkylbenzole sulfates and secondary alkane sulfonates.

Preferred nonionic solubilizers are selected from sorbitan fatty acidesters, polyalkoxylated fatty acid esters such as, for example,polyalkoxylated glycerides, polyalkoxylated sorbitan fatty acid estersand fatty acid esters of polyalkylene glycols, polyalkoxylated ethers offatty alcohols, tocopheryl compounds, and mixtures of two or morethereof. A fatty acid chain in these solubilizer compounds ordinarilycomprises 8 to 22 carbon atoms. Polyalkylene oxide blocks comprise onaverage 4 to 50 alkylene oxide units, preferably ethylene oxide units,per molecule.

Examples of suitable sorbitan fatty acid esters are sorbitanmonolaurate, sorbitan monopalmitate, sorbitan monostearate (e.g., Span™60), sorbitan monooleate (e.g., Span™ 80), sorbitan tristearate,sorbitan trioleate or sorbitan monolaurate.

Examples of suitable polyalkoxylated sorbitan fatty acid esters arepolyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitanmonopalmitate, polyoxyethylene (20) sorbitan monostearate,polyoxyethylene (20) sorbitan monooleate (e.g., Tween™ 80),polyoxyethylene (20) sorbitan tristearate (e.g., Tween™ 65),polyoxyethylene (20) sorbitan trioleate (e.g., Tween™ 85),polyoxyethylene (4) sorbitan monostearate, polyoxyethylene (4) sorbitanmonolaurate or polyoxyethylene (4) sorbitan monooleate.

Suitable polyalkoxylated glycerides are obtained for example byalkoxylation of natural or hydrogenated glycerides or bytransesterification of natural or hydrogenated glycerides withpolyalkylene glycols. Commercially available examples arepolyoxyethylene glycerol ricinoleate 35, polyoxyethylene glyceroltrihydroxystearate 40 (e.g., Cremophor RH™ 40 of BASF AG) andpolyalkoxylated glycerides including those obtainable under theproprietary names Gelucire™ and Labrafil™ from Gattefossé, e.g.,Gelucire™ 44/14 (lauroyl macrogol 32 glycerides prepared bytransesterification of hydrogenated palm kernel oil with PEG-1500),Gelucire™ 50/13 (stearoyl macrogol 32 glycerides, prepared bytransesterification of hydrogenated palm oil with PEG-1500) or Labrafil™M 1944 CS (oleoyl macrogol 6 glycerides prepared by transesterificationof apricot kernel oil with PEG-300).

A suitable fatty acid ester of polyalkylene glycols is, for example,PEG-660 hydroxystearic acid (polyglycol ester of 12-hydroxystearic acid(70 mol %) with 30 mol % ethylene glycol).

Suitable polyalkoxylated ethers of fatty alcohols are, for example, PEG(2) stearyl ether (e.g., Brij™ 72), macrogol 6 cetylstearyl ether ormacrogol 25 cetylstearyl ether.

In general, a tocopheryl compound useful herein corresponds to theformula

wherein Z is a linking group, R¹ and R² are, independently of oneanother, hydrogen or C₁-C₄ alkyl and n is an integer from 5 to 100,preferably 10 to 50. Typically, Z is the residue of an aliphatic dibasicacid such as glutaric, succinic or adipic acid. Preferably, both R¹ andR² are hydrogen.

A preferred tocopheryl compound is α-tocopheryl polyethylene glycolsuccinate, available for example as the proprietary product Vitamin ETPGS™. This is a water-soluble derivative of natural-source vitamin Eprepared by esterifying D-α-tocopheryl acid succinate with PEG-1000.

According to one preferred embodiment the pharmaceutically acceptablesolubilizer is selected from the group consisting of tocopherylcompounds having a polyalkylene glycol moiety (such as α-tocopherylpolyethylene glycol succinate), sorbitan fatty acid esters (such assorbitan monolaurate) and polyoxyethylene sorbitan fatty acid esters(such as polyoxyethylene sorbitan monolaurate) and combinations of twoor more thereof. This embodiment is particularly useful where the activeingredient is ABT-263 free base.

In another preferred embodiment the dosage form comprises at least onepharmaceutically acceptable nonionic solubilizer and at least onepharmaceutically acceptable anionic solubilizer. Preferably, thenonionic solubilizer is selected from the group consisting of sorbitanfatty acid esters, polyoxyethylene sorbitan fatty acid esters andα-tocopheryl polyethylene glycol succinate; and the anionic solubilizeris sodium lauryl sulfate (also referred to herein as SDS). Thisembodiment is particularly useful where the active ingredient is an acidaddition salt of ABT-263 such as ABT-263 bis-HCl.

Formation of a solid solution can be promoted by incorporating anon-volatile solvent for the active ingredient into the solid dispersionproduct. The non-volatile solvent is suitably selected from solventswith high dissolving power for a compound of Formula I, for exampleABT-263, which are liquid at ambient temperature and ambient pressure.

Nonlimiting examples of suitable solvents include liquid polyethyleneglycols, e.g., PEG-400; N-methylpyrrolidone;1,3-bis(pyrrolidon-1-yl)butane; and propylene glycol. A preferredsolvent is propylene glycol. The amount of the non-volatile solvent tobe used should not be so high as to compromise the mechanical propertiesof the solid dispersion product and usually is about 2% to about 10%,for example about 3% to about 5%, by weight of the solid dispersionproduct.

The pharmaceutically acceptable polymer may be selected fromwater-soluble polymers, water-dispersible polymers, water-swellablepolymers and mixtures thereof. Polymers are considered water-soluble ifthey form a clear homogeneous solution in water. When dissolved at 20°C. in an aqueous solution at 2% (w/v), the water-soluble polymerpreferably has an apparent viscosity of about 1 to about 5,000 mPa·s,more preferably about 1 to about 700 mPa·s, and most preferably about 5to about 100 mPa·s. Water-dispersible polymers are those that, whencontacted with water, form colloidal dispersions rather than a clearsolution. Upon contact with water or aqueous solutions, water-swellablepolymers typically form a rubbery gel.

Preferably, the pharmaceutically acceptable polymer employed incompositions of the present embodiment has a T_(g) of at least about 40°C., preferably at least about 50° C., most preferably about 80° C. toabout 180° C. The T_(g) value of a copolymer can be calculated as theweighted sum of the T_(g) values for homopolymers derived from each ofthe individual monomers, i, that make up the copolymer:T_(g)=ΣW_(i)X_(i) where W_(i) is the weight percent of monomer i in thecopolymer, and X, is the T_(g) value for the homopolymer derived frommonomer i. T_(g) values for homopolymers may be taken from Brandrup &Immergut, eds. (1975) Polymer Handbook, 2nd edition, John Wiley & Sons,Inc.

Various additives contained in the solid dispersion product or even theactive ingredient itself may exert a plasticizing effect on the polymerand thus depress the T_(g) of the polymer such that the final soliddispersion product has a somewhat lower T_(g) than the starting polymerused for its preparation. In general, the final solid dispersion producthas a T_(g) of 20° C. or higher, preferably 25° C. or higher, morepreferably 30° C. or higher and most preferably 40° C. or higher, e.g.,a T_(g) from about 45° C. to about 60° C.

For example, preferred pharmaceutically acceptable polymers can beselected from the group comprising homopolymers and copolymers ofN-vinyl lactams, especially homopolymers and copolymers of N-vinylpyrrolidone, e.g., polyvinylpyrrolidone (PVP), copolymers of N-vinylpyrrolidone and vinyl acetate or vinyl propionate, cellulose esters andcellulose ethers, in particular methylcellulose and ethylcellulose,hydroxyalkylcelluloses, in particular hydroxypropylcellulose,hydroxyalkylalkylcelluloses, in particularhydroxypropyl-methylcellulose, cellulose phthalates and succinates, inparticular cellulose acetate phthalate and hydroxypropylmethylcellulosephthalate, hydroxypropylmethylcellulose succinate andhydroxypropylmethylcellulose acetate succinate; high molecularpolyalkylene oxides such as polyethylene oxide and polypropylene oxideand copolymers of ethylene oxide and propylene oxide; polyvinylalcohol/polyethylene glycol graft copolymers (available as Kollicoat™ IRfrom BASF AG); polyacrylates and polymethacrylates such as methacrylicacid/ethyl acrylate copolymers, methacrylic acid/methyl methacrylatecopolymers, butyl methacrylate/2-dimethylaminoethyl methacrylatecopolymers, poly(hydroxyalkyl acrylates) and poly(hydroxyalkylmethacrylates); polyacrylamides; vinyl acetate polymers such ascopolymers of vinyl acetate and crotonic acid; partially hydrolyzedpolyvinyl acetate (also referred to as partially saponified “polyvinylalcohol”); polyvinyl alcohol; oligo- and polysaccharides such ascarrageenans, galactomannans and xanthan gum, and mixtures of two ormore thereof.

Among these, homopolymers or copolymers of N-vinyl pyrrolidone, inparticular a copolymer of N-vinyl pyrrolidone and vinyl acetate, arepreferred. A particularly preferred polymer is a copolymer of 60% byweight N-vinyl pyrrolidone and 40% by weight vinyl acetate.

A further polymer which can be suitably used is a mixture of PVP andpolyvinylacetate as sold, for example, under the proprietary nameKollidon® SR of BASF AG.

A solid dispersion product of the present embodiment may be prepared bya variety of methods.

Preferably, the solid dispersion product is prepared by melt-extrusion.Accordingly, the solid dispersion product is a melt-processed,solidified mixture. The melt-extrusion process comprises preparing ahomogeneous melt of an active ingredient or combination of activeingredients, the pharmaceutically acceptable polymer and thesolubilizer, and cooling the melt until it solidifies.

“Melting” in the present context means a transition into a liquid orrubbery state in which it is possible for one component to becomehomogeneously embedded in the other. Typically, one component will meltand the other components will dissolve in the melt, thus forming asolution. Melting usually involves heating above the softening point ofthe pharmaceutically acceptable polymer. Preparation of the melt cantake place in a variety of ways. Mixing of the components can take placebefore, during or after formation of the melt. For example, thecomponents can be mixed first and then melted, or they can besimultaneously mixed and melted. Usually, the melt is homogenized inorder to disperse the active ingredient efficiently. Also, it may beconvenient first to melt the pharmaceutically acceptable polymer andthen to admix and homogenize the active ingredient.

Usually, the melt temperature is in the range of about 70° C. to about250° C., preferably about 80° C. to about 180° C., and most preferablyabout 100° C. to about 140° C.

The active ingredient can be employed as such or as a solution ordispersion in a suitable solvent such as one or more alcohols, aliphatichydrocarbons or esters. Another solvent which can be used is liquidcarbon dioxide. The solvent is removed, e.g., evaporated, uponpreparation of the melt. Alternatively, solid dispersions of the activeingredient can be prepared with a non-volatile solvent for the activeingredient as previously mentioned.

Various additives may be included in the melt, for example flowregulators such as colloidal silica, lubricants, bulking agents(fillers), disintegrants, plasticizers, stabilizers such asantioxidants, light stabilizers, radical scavengers, or stabilizersagainst microbial attack.

The melting and/or mixing takes place in an apparatus customary for sucha purpose. Particularly suitable are extruders or kneaders. Suitableextruders include single screw extruders, intermeshing screw extrudersor multiscrew extruders, preferably twin-screw extruders, which can becorotating or counterrotating and, optionally, equipped with kneadingdisks or other screw elements for mixing or dispersing the melt. It willbe appreciated that the working temperatures will be determined by thekind of extruder or the kind of configuration within the extruder used.Part of the energy needed to melt, mix and dissolve the components inthe extruder can be provided by heating elements. However, the frictionand shearing of the material in the extruder may also provide asubstantial amount of energy to the mixture and aid in the formation ofa homogeneous melt of the components.

The extrudate exiting from the extruder ranges from pasty to viscous.Before allowing the extrudate to solidify, the extrudate may be directlyshaped into virtually any desired shape. Shaping of the extrudate may beconveniently carried out by a calendar with two counter-rotating rollerswith mutually matching depressions on their surface. A broad range oftablet forms can be attained by using rollers with different forms ofdepressions. If the rollers do not have depressions on their surface,films can be obtained. Alternatively, the extrudate is moulded into thedesired shape by injection-moulding. Alternatively, the extrudate issubjected to profile extrusion and cut into pieces, either before(hot-cut) or after solidification (cold-cut).

Additionally, foams can be formed if the extrudate contains a propellantsuch as a gas, e.g., carbon dioxide, or a volatile compound, e.g., a lowmolecular-weight hydrocarbon, or a compound that is thermallydecomposable to a gas. The propellant is dissolved in the extrudateunder the relatively high pressure conditions within the extruder and,when the extrudate emerges from the extruder die, the pressure issuddenly released. Thus the solvability of the propellant is decreasedand/or the propellant vaporizes so that a foam is formed.

Optionally, the resulting solid solution product is milled or ground togranules. The granules may then be filled into capsules or may becompacted. Compacting means a process whereby a powder mass comprisingthe granules is densified under high pressure in order to obtain acompact with low porosity, e.g., a tablet. Compression of the powdermass is usually done in a tablet press, more specifically in a steel diebetween two moving punches.

Preferably, the solid dosage form contains at least one additiveselected from flow regulators, disintegrants, bulking agents andlubricants.

At least one additive selected from flow regulators, disintegrants,bulking agents (fillers) and lubricants is preferably used in compactingthe granules. Disintegrants promote a rapid disintegration of thecompact in the stomach and help the liberated granules separate from oneanother. Suitable disintegrants are crosslinked polymers such ascrosslinked PVP (crospovidone) and crosslinked sodiumcarboxymethylcellulose. Suitable bulking agents (also referred to as“fillers”) can be selected from mannitol, lactose, calcium hydrogenphosphate, microcrystalline cellulose (e.g., Avicel™), magnesium oxide,potato and corn starches, isomalt and polyvinyl alcohol. Suitable flowregulators can be selected from highly dispersed silica (e.g., Aerosil™)(also referred to as colloidal silicon dioxide), and animal andvegetable fats and waxes. A lubricant is preferably used in compactingthe granules. Suitable lubricants can be selected from polyethyleneglycol (e.g., having a molecular weight of about 1,000 to about 6,000),magnesium and calcium stearates, sodium stearyl fumarate, talc, and thelike.

Various other additives may be used, for example dyes such as azo dyes,organic or inorganic pigments such as aluminum oxide or titaniumdioxide, or dyes of natural origin; stabilizers such as antioxidants,light stabilizers, radical scavengers, or stabilizers against microbialattack. Such additives are known to those skilled in the art, andnon-limiting examples include Vitamin E and derivatives thereof (e.g.,Vitamin E-TPGST™), butylhydroxytoluene (BTH), cysteine, and ascorbicacid and derivatives thereof.

Dosage forms according to the present embodiment may consist of severallayers, as for example in laminated or multilayer tablets. They can bein open or closed form. “Closed dosage forms” are those in which onelayer is completely surrounded by at least one other layer. Multilayerforms have the advantage that two active ingredients which areincompatible with one another can be processed, or that the releasecharacteristics of the active ingredient(s) can be controlled. Forexample, it is possible to provide an initial dose by including anactive ingredient in an outer layer, and a maintenance dose by includingthe active ingredient in an inner layer. Multilayer tablet types may beproduced by compressing two or more layers of granules. Alternatively,multilayer dosage forms may be produced by a process known as“coextrusion”. In essence, the process comprises preparation of at leasttwo different melt compositions as explained above, and passing thesemolten compositions into a joint coextrusion die. The shape of thecoextrusion die depends on the required drug form. For example, dieswith a plain die gap, called slot dies, and dies with an annular slitare suitable.

In order to facilitate oral administration of such a dosage form, it isadvantageous to give the dosage form an appropriate shape. Large tabletsare therefore preferably elongated rather than round in shape, tofacilitate comfortable swallowing.

An optional film coat on the tablet further contributes to ease ofswallowing. A film coat also improves taste and provides an elegantappearance. If desired, the film coat may be an enteric coat. The filmcoat usually includes a polymeric film-forming material such ashydroxypropylmethylcellulose, hydroxypropylcellulose, or an acrylate ormethacrylate copolymer. Besides a film-forming polymer, the film coatmay further comprise a plasticizer, e.g., polyethylene glycol, asurfactant, e.g., a polyoxyethylene sorbitan ester, and optionally apigment, e.g., titanium dioxide or iron oxide. The film coat may alsocomprise talc as an anti-adherent. The film coat if present usuallyaccounts for less than about 5% by weight of the dosage form.

In an alternative process for preparing a solid dosage form, the soliddispersion product is ground and filled into a capsule shell. Suitablematerials for capsule shells are known in the art, and include forexample gelatin, gums such as carrageenan or gellan, and cellulose orcellulose derivatives such as hydroxypropylmethylcellulose.

It has been found that a solid dispersion of ABT-263 according to thepresent embodiment not only shows adequate bioavailability after oraladministration but also results in a storage-stable, ready-to-use dosageform. Quite surprisingly, in such a solid dispersion the ABT-263molecule, despite its essentially non-crystalline amorphous state, islargely resistant against oxidation even in presence of only a minoramount of antioxidant or absence of any antioxidant.

However, optionally an HCA, for example a sulfur-containing antioxidant,can be included in the composition of the present embodiment if sodesired.

Sixth Composition Embodiment

A composition of the sixth embodiment set forth hereinabove comprises(a) a compound of Formula I or a pharmaceutically acceptable saltthereof, for example ABT-263 free base or ABT-263 bis-HCl, in solidparticulate form and in a free base equivalent amount of at least about2.5% by weight of the composition, and (b) a plurality ofpharmaceutically acceptable excipients including at least a soliddiluent and a solid disintegrant.

Illustratively, the active ingredient concentration in a composition ofthe present embodiment is at least about 5%, at least about 10%, atleast about 15%, at least about 20%, at least about 25% or at leastabout 30% by weight of the formulation, and can be as high as 40% byweight or, in some instances, even higher.

It is generally preferred that the solid particulate form of the activeingredient used in the composition should be a crystalline form. In thecase of ABT-263, the product prepared by the process described in the'135 publication is non-crystalline and is generally unsuitable forformulation as a solid dosage form of the present embodiment. For thisreason, the composition preferably contains as API a crystalline form ofthe free base, e.g., ABT-263 free base crystalline Form I or Form II asdescribed hereinabove, or a crystalline salt, such as ABT-263 bis-HCl.

Particle size of the API is not narrowly critical, though resultssuggest that reduction in particle size can improve bioavailability. Incompositions of the invention, the D₉₀ particle size (90% by volume ofthe API particles in their longest dimension are smaller than this) istypically about 2.5 to about 50 μm, for example about 3 to about 30 μm.API in the upper part of this D₉₀ range is typically unmilled. Reductionin particle size to the lower part of the D₉₀ range is achievable, forexample, by pin-milling or jet-milling. In some compositions, unmilledAPI having a D₉₀ of about 20 to about 30 μm is used. In othercompositions, pin-milled or jet-milled API having a D₉₀ of about 3 toabout 10 μm is used. In still other compositions, API of intermediateD₉₀, for example about 10 to about 20 μm, is used.

A composition of the present embodiment comprises, in addition to theAPI, a plurality of pharmaceutically acceptable excipients including atleast one or more solid diluents and one or more solid disintegrants.Optionally, the excipients further include one or more binding agents,wetting agents and/or antifrictional agents (lubricants, anti-adherentsand/or glidants). Many excipients have two or more functions in apharmaceutical composition. Characterization herein of a particularexcipient as having a certain function, e.g., diluent, disintegrant,binding agent, etc., should not be read as limiting to that function.Further information on excipients can be found in standard referenceworks such as Kibbe, ed. (2000) Handbook of Pharmaceutical Excipients,3rd edition, Washington: American Pharmaceutical Association).

Suitable diluents illustratively include, either individually or incombination, lactose, including anhydrous lactose and lactosemonohydrate; lactitol; maltitol; mannitol; sorbitol; xylitol; dextroseand dextrose monohydrate; fructose; sucrose and sucrose-based diluentssuch as compressible sugar, confectioner's sugar and sugar spheres;maltose; inositol; hydrolyzed cereal solids; starches (e.g., cornstarch, wheat starch, rice starch, potato starch, tapioca starch, etc.),starch components such as amylose and dextrates, and modified orprocessed starches such as pregelatinized starch; dextrins; cellulosesincluding powdered cellulose, microcrystalline cellulose, silicifiedmicrocrystalline cellulose, food grade sources of α- and amorphouscellulose and powdered cellulose, and cellulose acetate; calcium saltsincluding calcium carbonate, tribasic calcium phosphate, dibasic calciumphosphate dihydrate, monobasic calcium sulfate monohydrate, calciumsulfate and granular calcium lactate trihydrate; magnesium carbonate;magnesium oxide; bentonite; kaolin; sodium chloride; and the like. Suchdiluents, if present, typically constitute in total about 5% to about95%, for example about 20% to about 90%, or about 50% to about 85%, byweight of the composition. The diluent or diluents selected preferablyexhibit suitable flow properties and, where tablets are desired,compressibility.

Microcrystalline cellulose and silicified microcrystalline cellulose areparticularly useful diluents, and are optionally used in combinationwith a water-soluble diluent such as mannitol. Illustratively, asuitable weight ratio of microcrystalline cellulose or silicifiedmicrocrystalline cellulose to mannitol is about 10:1 to about 1:1, butratios outside this range can be useful in particular circumstances.

Suitable disintegrants include, either individually or in combination,starches including pregelatinized starch and sodium starch glycolate;clays; magnesium aluminum silicate; cellulose-based disintegrants suchas powdered cellulose, microcrystalline cellulose, methylcellulose,low-substituted hydroxypropylcellulose, carmellose, carmellose calcium,carmellose sodium and croscarmellose sodium; alginates; povidone;crospovidone; polacrilin potassium; gums such as agar, guar, locustbean, karaya, pectin and tragacanth gums; colloidal silicon dioxide; andthe like. One or more disintegrants, if present, typically constitute intotal about 0.2% to about 30%, for example about 0.5% to about 20%, orabout 1% to about 10%, by weight of the composition.

Sodium starch glycolate is a particularly useful disintegrant, andtypically constitutes in total about 1% to about 20%, for example about2% to about 15%, or about 5% to about 10%, by weight of the composition.

Binding agents or adhesives are useful excipients, particularly wherethe composition is in the form of a tablet. Such binding agents andadhesives should impart sufficient cohesion to the blend being tabletedto allow for normal processing operations such as sizing, lubrication,compression and packaging, but still allow the tablet to disintegrateand the composition to be absorbed upon ingestion. Suitable bindingagents and adhesives include, either individually or in combination,acacia; tragacanth; glucose; polydextrose; starch includingpregelatinized starch; gelatin; modified celluloses includingmethylcellulose, carmellose sodium, hydroxypropylmethylcellulose (HPMC),hydroxypropylcellulose, hydroxyethylcellulose and ethylcellulose;dextrins including maltodextrin; zein; alginic acid and salts of alginicacid, for example sodium alginate; magnesium aluminum silicate;bentonite; polyethylene glycol (PEG); polyethylene oxide; guar gum;polysaccharide acids; polyvinylpyrrolidone (povidone or PVP), forexample povidone K-15, K-30 and K-29/32; polyacrylic acids (carbomers);polymethacrylates; and the like. One or more binding agents and/oradhesives, if present, typically constitute in total about 0.5% to about25%, for example about 1% to about 15%, or about 1.5% to about 10%, byweight of the composition.

Povidone and hydroxypropylcellulose, either individually or incombination, are particularly useful binding agents for tabletformulations, and, if present, typically constitute about 0.5% to about15%, for example about 1% to about 10%, or about 2% to about 8%, byweight of the composition.

Wetting agents, if present, are normally selected to maintain the drugin close association with water, a condition that can improvebioavailability of the composition. Non-limiting examples of surfactantsthat can be used as wetting agents include, either individually or incombination, quaternary ammonium compounds, for example benzalkoniumchloride, benzethonium chloride and cetylpyridinium chloride; dioctylsodium sulfosuccinate; polyoxyethylene alkylphenyl ethers, for examplenonoxynol 9, nonoxynol 10 and octoxynol 9; poloxamers (polyoxyethyleneand polyoxypropylene block copolymers); polyoxyethylene fatty acidglycerides and oils, for example polyoxyethylene (8) caprylic/capricmono- and diglycerides, polyoxyethylene (35) castor oil andpolyoxyethylene (40) hydrogenated castor oil; polyoxyethylene alkylethers, for example ceteth-10, laureth-4, laureth-23, oleth-2, oleth-10,oleth-20, steareth-2, steareth-10, steareth-20, steareth-100 andpolyoxyethylene (20) cetostearyl ether; polyoxyethylene fatty acidesters, for example polyoxyethylene (20) stearate, polyoxyethylene (40)stearate and polyoxyethylene (100) stearate; sorbitan esters, forexample sorbitan monolaurate, sorbitan monooleate, sorbitanmonopalmitate and sorbitan monostearate; polyoxyethylene sorbitanesters, for example polysorbate 20 and polysorbate 80; propylene glycolfatty acid esters, for example propylene glycol laurate; sodium laurylsulfate; fatty acids and salts thereof, for example oleic acid, sodiumoleate and triethanolamine oleate; glyceryl fatty acid esters, forexample glyceryl monooleate, glyceryl monostearate and glycerylpalmitostearate; α-tocopherol polyethylene glycol (1000) succinate(TPGS); tyloxapol; and the like. One or more wetting agents, if present,typically constitute in total about 0.1% to about 15%, for example about0.2% to about 10%, or about 0.5% to about 7%, by weight of thecomposition.

Nonionic surfactants, more particularly poloxamers, are examples ofwetting agents that can be useful herein. Illustratively, a poloxamersuch as Pluronic™ F127, if present, can constitute about 0.1% to about10%, for example about 0.2% to about 7%, or about 0.5% to about 5%, byweight of the composition.

Lubricants reduce friction between a tableting mixture and tabletingequipment during compression of tablet formulations. Suitable lubricantsinclude, either individually or in combination, glyceryl behenate;stearic acid and salts thereof, including magnesium, calcium and sodiumstearates; hydrogenated vegetable oils; glyceryl palmitostearate; talc;waxes; sodium benzoate; sodium acetate; sodium fumarate; sodium stearylfumarate; PEGs (e.g., PEG 4000 and PEG 6000); poloxamers; polyvinylalcohol; sodium oleate; sodium lauryl sulfate; magnesium lauryl sulfate;and the like. One or more lubricants, if present, typically constitutein total about 0.05% to about 10%, for example about 0.1% to about 5%,or about 0.2% to about 2%, by weight of the composition. Sodium stearylfumarate is a particularly useful lubricant.

Anti-adherents reduce sticking of a tablet formulation to equipmentsurfaces. Suitable anti-adherents include, either individually or incombination, talc, colloidal silicon dioxide, starch, DL-leucine, sodiumlauryl sulfate and metallic stearates. One or more anti-adherents, ifpresent, typically constitute in total about 0.05% to about 10%, forexample about 0.1% to about 7%, or about 0.2% to about 5%, by weight ofthe composition. Colloidal silicon dioxide is a particularly usefulanti-adherent.

Glidants improve flow properties and reduce static in a tabletingmixture. Suitable glidants include, either individually or incombination, colloidal silicon dioxide, starch, powdered cellulose,sodium lauryl sulfate, magnesium trisilicate and metallic stearates. Oneor more glidants, if present, typically constitute in total about 0.05%to about 10%, for example about 0.1% to about 7%, or about 0.2% to about5%, by weight of the composition. Colloidal silicon dioxide is aparticularly useful glidant.

Other excipients such as buffering agents, stabilizers, antioxidants,antimicrobials, colorants, flavors and sweeteners are known in thepharmaceutical art and can be used in compositions of the presentinvention. Tablets can be uncoated or can comprise a core that iscoated, for example with a nonfunctional film or a release-modifying orenteric coating. Capsules can have hard or soft shells comprising, forexample, gelatin (in the form of hard gelatin capsules or soft elasticgelatin capsules), starch, carrageenan and/or HPMC, optionally togetherwith one or more plasticizers.

Solid dosage forms according to the present embodiment not only showadequate bioavailability after oral administration but exhibitacceptable storage-stability, being relatively resistant to oxidativedegradation of the active ingredient even in presence of only a minoramount of antioxidant or absence of any antioxidant.

However, optionally an HCA, for example a sulfur-containing antioxidant,can be included in the composition of the present embodiment if sodesired.

Any suitable process of pharmacy can be used to prepare a composition ofthe present embodiment, including dry blending with or without directcompression, and wet or dry granulation. In the illustrative,non-limiting processes and compositions shown below, API can be used inunmilled form, e.g., with a D₉₀ particle size of about 20 to about 30μm, or after milling to a desired size, e.g., pin-milled or jet-milledto a D₉₀ particle size of about 3 to about 10 μm.

An illustrative dry blending process is as follows. API (e.g., ABT-263bis-HCl) is mixed with excipients except lubricant, for example byblending in a V-blender for approximately 20 minutes. Lubricant is thenadded. The resulting powder blend is compressed, for example at 500 lb,in a tablet press with suitable tooling to provide the size and shape oftablets desired. Alternatively, the powder blend is filled intocapsules.

An illustrative composition prepared by the above process consists ofthe following ingredients (all percentages by weight):

ABT-263 bis-HCl 10.75% (10% free base equivalent) silicifiedmicrocrystalline cellulose 49.00% mannitol 20.00% pregelatinized starch5.00% sodium starch glycolate 10.00% poloxamer (Pluronic ™ F127) 4.00%colloidal silicon dioxide 1.00% sodium stearyl fumarate 0.25%Tablets of 50 mg ABT-263 dosage strength (total tablet weight 500 mg)are prepared from the above ingredients in a Carver press at 500 lb,with round tooling.

A first illustrative wet granulation process is as follows. API (e.g.,ABT-263 bis-HCl) is suspended in a binder/surfactant solution(granulation liquid), then added to a blend of diluent(s) anddisintegrant(s) in a food processor to prepare a granulate.

A second illustrative wet granulation process is as follows. API (e.g.,ABT-263 bis-HCl) is mixed with excipients, including granulation liquidbut excluding lubricant, and granulated in a food processor. Thegranules are dried and passed through a 20 mesh screen. Lubricant isthen added.

A third illustrative wet granulation process is as follows. API (e.g.,ABT-263 bis-HCl) is mixed with excipients, including granulation liquidand a first amount of disintegrant (intragranular excipients) butexcluding lubricant, and granulated in a food processor. The granulesare dried and passed through a 20 mesh screen. A second amount ofdisintegrant, lubricant and optionally other extragranular excipient(s)are then added.

Granules prepared by any of the above wet granulation processes can becompressed, for example at 500 lb, in a tablet press with suitabletooling to provide the size and shape of tablets desired. Alternatively,the granules can be filled into capsules.

A first illustrative tablet composition that can be prepared by any ofthe above wet granulation processes consists of the followingingredients (all percentages by weight):

ABT-263 bis-HCl 10.75%  (10% free base equivalent) microcrystallinecellulose 83.50%  povidone K-30 3.00% crospovidone 1.50% poloxamer(Pluronic ™ F127) 1.00% sodium stearyl fumarate 0.25%

A second illustrative tablet composition that can be prepared by any ofthe above wet granulation processes consists of the followingingredients (all percentages by weight):

ABT-263 bis-HCl 5.38% (5% free base equivalent) microcrystallinecellulose 85.87%  povidone K-30 3.00% crospovidone 1.50% poloxamer(Pluronic ™ F127) 4.00% sodium stearyl fumarate 0.25%

A third illustrative tablet composition that can be prepared by any ofthe above wet granulation processes consists of the followingingredients (all percentages by weight):

ABT-263 bis-HCl 10.75% (10% free base equivalent) microcrystallinecellulose 50.00% mannitol 20.00% povidone K-30 5.00% sodium starchglycolate 10.00% poloxamer (Pluronic ™ F127) 4.00% sodium stearylfumarate 0.25%

Tablets containing a 50 mg dose of ABT-263 are prepared from any of theabove wet granulations.

An illustrative capsule composition that can be prepared by any of theabove wet granulation processes consists of the following ingredients(all percentages by weight):

ABT-263 bis-HCl 10.75% (10% free base equivalent) microcrystallinecellulose 50.00% mannitol 30.00% hydroxypropylcellulose 3.00% sodiumstarch glycolate 5.00% poloxamer (Pluronic ™ F127) 1.00% sodium stearylfumarate 0.25%The composition is filled into size 0 capsules.

Bioavailability and Administration

In any of the above embodiments, and others not fully described hereinbut evident to the ordinarily skilled reader of the presentspecification, the formulation ingredients and amounts thereof can beselected to provide enhanced bioabsorption by comparison with a standardsolution of the drug when administered orally. Such enhancedbioabsorption versus the standard solution can be evidenced, forexample, by a pharmacokinetic (PK) profile having one or more of ahigher C_(max) or an increased bioavailability as measured by AUC, forexample AUC₀₋₂₄ or AUC_(0-∞). Illustratively, bioavailability can beexpressed as a percentage, for example using the parameter F, whichcomputes AUC for oral delivery of a test composition as a percentage ofAUC for intravenous (i.v.) delivery of the drug in a suitable solvent,taking into account any difference between oral and i.v. doses.

The standard solution in the case of ABT-263 can be, for example, asolution of ABT-263 free base in a carrier consisting of 10% DMSO inPEG-400, or a formulation referenced herein as “Formulation C”, which isa solution of ABT-263 bis-HCl solution at a free base equivalentconcentration of 25 mg/ml in a carrier liquid consisting of 90%phosphatidylcholine+medium chain triglycerides 53/29 and 10% dehydratedalcohol USP (meeting standards set forth in the United StatesPharmacopeia).

Bioavailability can be determined by PK studies in humans or in anysuitable model species. For present purposes, a dog model is generallysuitable. In various illustrative embodiments, where the drug is ABT-263or a salt thereof, compositions of the invention exhibit oralbioavailability of at least about 15%, at least about 30%, at leastabout 35% or at least about 40%, up to or exceeding about 50%, in a dogmodel, when administered as a single dose of about 2.5 to about 10 mg/kgto fasting or non-fasting animals.

In one example, the composition comprises ABT-263 or a salt thereof anda carrier comprising ingredients and amounts thereof selected to providea PK profile upon oral administration of the composition in anon-fasting dog model exhibiting a bioavailability of at least about15%.

In one example, the composition comprises ABT-263 or a salt thereof anda carrier comprising ingredients and amounts thereof selected to providea PK profile upon oral administration of the composition in anon-fasting dog model exhibiting a bioavailability of at least about30%.

In one example, the composition comprises ABT-263 or a salt thereof anda carrier comprising ingredients and amounts thereof selected to providea PK profile upon oral administration of the composition in anon-fasting dog model exhibiting a bioavailability of at least about40%.

The potential of the present invention to provide bioavailability, forexample of ABT-263, substantially greater, for example at least about1.5× or at least about 2× greater, than that of the solution in 10% DMSOin PEG-400 described in above-cited U.S. Patent Application PublicationNo. 2007/0027135, is an unexpected benefit of great practical value,especially in view of the fact that formulation changes apparently havelittle effect on bioavailability of earlier generations of Bcl-2 proteinfamily inhibitors such as ABT-737. Bioavailability in a rat model ofABT-737, formulated in 90% phosphatidylcholine+medium chaintriglycerides 53/29 and 10% ethanol, was only 3.3%, not markedlydifferent from that of other formulations tested.

Sufficient bioavailability of an ABT-263 composition is evidenced insome embodiments by one or both of

(a) an ABT-263 AUC₀₋₂₄ of at least about 20 μg·h/ml, and/or

(b) an ABT-263 C_(max) of at least about 2.5 μg/ml,

in a single-dose non-fasting human PK study at an ABT-263 free baseequivalent dose of about 200 to about 400 mg.

Sufficient bioavailability of an ABT-263 composition is evidenced inother embodiments by a steady-state ABT-263 C_(am), of about 1 to about5 μg/ml and a steady-state ABT-263 C_(max) of about 3 to about 8 μg/mlin a non-fasting human pharmacokinetic study at a daily ABT-263 freebase equivalent dose of about 200 to about 400 mg.

In particular embodiments, an ABT-263 composition is at leastsubstantially bioequivalent to Formulation C as defined above.

The term “substantially bioequivalent” herein means exhibiting, in ahuman PK single- or multiple-dose study in fasting or non-fastingconditions, substantially equal C_(max) and substantially equal exposuremeasured as AUC, for example AUC₀₋₂₄, AUC₀₋₄₈ or AUC_(0-∞). Thecompositions being compared for substantial bioequivalence should beadministered at the same dose or doses, expressed as free baseequivalent. If a multiple-dose study is used to draw the comparison, itis the steady-state values of C_(max) and AUC that are used. In thepresent context, C_(max) or AUC of a test composition is “substantiallyequal” if it is no less than 80% and no greater than 125% of thecorresponding parameter in a reference composition (e.g., FormulationC).

Compositions embraced herein, including compositions described generallyor with specificity herein, are useful for orally delivering a compoundof Formula I, for example ABT-263, or a pharmaceutically acceptable saltthereof, to a subject. Accordingly, a method of the invention fordelivering a compound of Formula I, for example ABT-263, or apharmaceutically acceptable salt thereof, to a subject comprises orallyadministering a composition as described above.

The subject can be human or non-human (e.g., a farm, zoo, work orcompanion animal, or a laboratory animal used as a model) but in animportant embodiment the subject is a human patient in need of the drug,for example to treat a disease characterized by apoptotic dysfunctionand/or overexpression of an anti-apoptotic Bcl-2 family protein. A humansubject can be male or female and of any age. The patient is typicallyan adult, but a method of the invention can be useful to treat achildhood cancer such as leukemia, for example acute lymphocyticleukemia, in a pediatric patient.

The composition is normally administered in an amount providing atherapeutically effective daily dose of the drug. The term “daily dose”herein means the amount of drug administered per day, regardless of thefrequency of administration. For example, if the subject receives a unitdose of 150 mg twice daily, the daily dose is 300 mg. Use of the term“daily dose” will be understood not to imply that the specified dosageamount is necessarily administered once daily. However, in a particularembodiment the dosing frequency is once daily (q.d.), and the daily doseand unit dose are in this embodiment the same thing.

What constitutes a therapeutically effective dose depends on thebioavailability of the particular formulation, the subject (includingspecies and body weight of the subject), the disease (e.g., theparticular type of cancer) to be treated, the stage and/or severity ofthe disease, the individual subject's tolerance of the compound, whetherthe compound is administered in monotherapy or in combination with oneor more other drugs, e.g., other chemotherapeutics for treatment ofcancer, and other factors. Thus the daily dose can vary within widemargins, for example from about 10 to about 1,000 mg. Greater or lesserdaily doses can be appropriate in specific situations. It will beunderstood that recitation herein of a “therapeutically effective” doseherein does not necessarily require that the drug be therapeuticallyeffective if only a single such dose is administered; typicallytherapeutic efficacy depends on the composition being administeredrepeatedly according to a regimen involving appropriate frequency andduration of administration. It is strongly preferred that, while thedaily dose selected is sufficient to provide benefit in terms oftreating the cancer, it should not be sufficient to provoke an adverseside-effect to an unacceptable or intolerable degree. A suitabletherapeutically effective dose can be selected by the physician ofordinary skill without undue experimentation based on the disclosureherein and on art cited herein, taking into account factors such asthose mentioned above. The physician may, for example, start a cancerpatient on a course of therapy with a relatively low daily dose andtitrate the dose upwards over a period of days or weeks, to reduce riskof adverse side-effects.

Illustratively, suitable doses of ABT-263 are generally about 25 toabout 1,000 mg/day, more typically about 50 to about 500 mg/day or about200 to about 400 mg/day, for example about 50, about 100, about 150,about 200, about 250, about 300, about 350, about 400, about 450 orabout 500 mg/day, administered at an average dosage interval of about 3hours to about 7 days, for example about 8 hours to about 3 days, orabout 12 hours to about 2 days. In most cases a once-daily (q.d.)administration regimen is suitable.

An “average dosage interval” herein is defined as a span of time, forexample one day or one week, divided by the number of unit dosesadministered over that span of time. For example, where a drug isadministered three times a day, around 8 am, around noon and around 6μm, the average dosage interval is 8 hours (a 24-hour time span dividedby 3). If the drug is formulated as a discrete dosage form such as atablet or capsule, a plurality (e.g., 2 to 4) of dosage formsadministered at one time is considered a unit dose for the purpose ofdefining the average dosage interval.

A daily dosage amount and dosage interval can, in some embodiments, beselected to maintain a plasma concentration of ABT-263 in a range ofabout 0.5 to about 10 μg/ml. Thus, during a course of ABT-263 therapyaccording to such embodiments, the steady-state peak plasmaconcentration (C_(max)) should in general not exceed about 10 μg/ml, andthe steady-state trough plasma concentration (C_(min)) should in generalnot fall below about 0.5 μg/ml. It will further be found desirable toselect, within the ranges provided above, a daily dosage amount andaverage dosage interval effective to provide a C_(max)/C_(mm) ratio notgreater than about 5, for example not greater than about 3, atsteady-state. It will be understood that longer dosage intervals willtend to result in greater C_(max)/C_(mm) ratios. Illustratively, atsteady-state, an ABT-263 C_(max) of about 3 to about 8 μg/ml and C_(min)of about 1 to about 5 μg/ml can be targeted by the present method.Steady-state values of C_(max) and C_(mm) can be established in a humanPK study, for example conducted according to standard protocolsincluding but not limited to those acceptable to a regulatory agencysuch as the U.S. Food and Drug Administration (FDA).

In the case of solid unit dosage forms, one to a small plurality oftablets or capsules can be swallowed whole, typically with the aid ofwater or other imbibable liquid to help the swallowing process.Optionally, tablets may be broken before swallowing and can be scored tofacilitate even breakage.

As compositions of the present invention are believed to exhibit only aminor food effect, administration according to the present embodimentcan be with or without food, i.e., in a non-fasting or fastingcondition. It is generally preferred to administer the presentcompositions to a non-fasting patient.

Method for Treating Disease

In still further embodiments of the invention, there is provided amethod for treating a disease characterized by apoptotic dysfunctionand/or overexpression of an anti-apoptotic Bcl-2 family protein,comprising administering to a subject having the disease atherapeutically effective amount of a compound of Formula I, for exampleABT-263, or a pharmaceutically acceptable salt thereof, formulated in acomposition as described herein.

Formulations of the present invention are suitable for use inmonotherapy or in combination therapy, for example with otherchemotherapeutics or with ionizing radiation. A particular advantage ofthe present invention is that it permits once-daily oral administration,a regimen which is convenient for the patient who is undergoingtreatment with other orally administered drugs on a once-daily regimen.Oral administration is easily accomplished by the patient him/herself orby a caregiver in the patient's home; it is also a convenient route ofadministration for patients in a hospital or residential care setting.

Combination therapies illustratively include administration of acomposition of the invention, for example such a composition comprisingABT-263, concomitantly with one or more of bortezomid, carboplatin,cisplatin, cyclophosphamide, dacarbazine, dexamethasone, docetaxel,doxorubicin, etoposide, fludarabine, hydroxydoxorubicin, irinotecan,paclitaxel, rapamycin, rituximab, vincristine and the like, for examplewith a polytherapy such as CHOP(cyclophosphamide+hydroxydoxorubicin+vincristine+prednisone), RCVP(rituximab+cyclophosphamide+vincristine+prednisone), R-CHOP(rituximab+CHOP) or DA-EPOCH-R dose-adjusted etoposide, prednisone,vincristine, cyclophosphamide, doxorubicin and rituximab).

A composition of the invention, for example such a compositioncomprising ABT-263, can be administered in combination therapy with oneor more therapeutic agents that include, but are not limited to,angiogenesis inhibitors, antiproliferative agents, other apoptosispromoters (for example, Bcl-xL, Bcl-w and Bfl-1 inhibitors), activatorsof a death receptor pathway, BiTE (bi-specific T-cell engager)antibodies, dual variable domain binding proteins (DVDs), inhibitors ofapoptosis proteins (IAPs), microRNAs, mitogen-activated extracellularsignal-regulated kinase inhibitors, multivalent binding proteins,poly-ADP (adenosine diphosphate)-ribose polymerase (PARP) inhibitors,small inhibitory ribonucleic acids (siRNAs), kinase inhibitors, receptortyrosine kinase inhibitors, aurora kinase inhibitors, polo-like kinaseinhibitors, bcr-abl kinase inhibitors, growth factor inhibitors, COX-2inhibitors, non-steroidal anti-inflammatory drugs (NSAIDs), antimitoticagents, alkylating agents, antimetabolites, intercalating antibiotics,platinum-containing chemotherapeutic agents, growth factor inhibitors,ionizing radiation, cell cycle inhibitors, enzymes, topoisomeraseinhibitors, biologic response modifiers, immunologicals, antibodies,hormonal therapies, retinoids, deltoids, plant alkaloids, proteasomeinhibitors, HSP-90 inhibitors, histone deacetylase (HDAC) inhibitors,purine analogs, pyrimidine analogs, MEK inhibitors, CDK inhibitors,ErbB2 receptor inhibitors, mTOR inhibitors as well as other antitumoragents.

Angiogenesis inhibitors include, but are not limited to, EGFRinhibitors, PDGFR inhibitors, VEGFR inhibitors, TIE2 inhibitors, IGF1Rinhibitors, matrix metalloproteinase 2 (MMP-2) inhibitors, matrixmetalloproteinase 9 (MMP-9) inhibitors and thrombospondin analogs.

Examples of EGFR inhibitors include, but are not limited to, gefitinib,erlotinib, cetuximab, EMD-7200, ABX-EGF, HR3, IgA antibodies, TP-38(IVAX), EGFR fusion protein, EGF-vaccine, anti-EGFR immunoliposomes andlapatinib.

Examples of PDGFR inhibitors include, but are not limited to, CP-673451and CP-868596.

Examples of VEGFR inhibitors include, but are not limited to,bevacizumab, sunitinib, sorafenib, CP-547632, axitinib, vandetanib,AEE788, AZD-2171, VEGF trap, vatalanib, pegaptanib, IM862, pazopanib,ABT-869 and angiozyme.

Bcl-2 family protein inhibitors other than ABT-263 include, but are notlimited to, AT-101 ((−)gossypol), Genasense™ Bcl-2-targeting antisenseoligonucleotide (G3139 or oblimersen), IPI-194, IPI-565, ABT-737, GX-070(obatoclax) and the like.

Activators of a death receptor pathway include, but are not limited to,TRAIL, antibodies or other agents that target death receptors (e.g., DR4and DR5) such as apomab, conatumumab, ETR2-ST01, GDC0145 (lexatumumab),HGS-1029, LBY-135, PRO-1762 and trastuzumab.

Examples of thrombospondin analogs include, but are not limited to,TSP-1, ABT-510, ABT-567 and ABT-898.

Examples of aurora kinase inhibitors include, but are not limited to,VX-680, AZD-1152 and MLN-8054.

An example of a polo-like kinase inhibitor includes, but is not limitedto, BI-2536.

Examples of bcr-abl kinase inhibitors include, but are not limited to,imatinib and dasatinib.

Examples of platinum-containing agents include, but are not limited to,cisplatin, carboplatin, eptaplatin, lobaplatin, nedaplatin, oxaliplatinand satraplatin.

Examples of mTOR inhibitors include, but are not limited to, CCI-779,rapamycin, temsirolimus, everolimus, RAD001 and AP-23573.

Examples of HSP-90 inhibitors include, but are not limited to,geldanamycin, radicicol, 17-AAG, KOS-953, 17-DMAG, CNF-101, CNF-1010,17-AAG-nab, NCS-683664, efungumab, CNF-2024, PU3, PU24FC1, VER-49009,IPI-504, SNX-2112 and STA-9090.

Examples of HDAC inhibitors include, but are not limited to,suberoylanilide hydroxamic acid (SAHA), MS-275, valproic acid, TSA,LAQ-824, trapoxin and depsipeptide.

Examples of MEK inhibitors include, but are not limited to, PD-325901,ARRY-142886, ARRY-438162 and PD-98059.

Examples of CDK inhibitors include, but are not limited to,flavopyridol, MCS-5A, CVT-2584, seliciclib ZK-304709, PHA-690509,BMI-1040, GPC-286199, BMS-387032, PD-332991 and AZD-5438.

Examples of COX-2 inhibitors include, but are not limited to, celecoxib,parecoxib, deracoxib, ABT-963, etoricoxib, lumiracoxib, BMS-347070, RS57067, NS-398, valdecoxib, rofecoxib, SD-8381,4-methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoylphenyl)-1H-pyrrole, T-614,JTE-522, S-2474, SVT-2016, CT-3 and SC-58125.

Examples of NSAIDs include, but are not limited to, salsalate,diflunisal, ibuprofen, ketoprofen, nabumetone, piroxicam, naproxen,diclofenac, indomethacin, sulindac, tolmetin, etodolac, ketorolac andoxaprozin.

Examples of ErbB2 receptor inhibitors include, but are not limited to,CP-724714, canertinib, trastuzumab, petuzumab, TAK-165, ionafamib,GW-282974, EKB-569, PI-166, dHER2, APC-8024, anti-HER/2neu bispecificantibody B7.her2IgG3 and HER2 trifunctional bispecific antibodies mABAR-209 and mAB 2B-1.

Examples of alkylating agents include, but are not limited to, nitrogenmustard N-oxide, cyclophosphamide, ifosfamide, trofosfamide,chlorambucil, melphalan, busulfan, mitobronitol, carboquone, thiotepa,ranimustine, nimustine, Cloretazine™ (laromustine), AMD-473,altretamine, AP-5280, apaziquone, brostallicin, bendamustine,carmustine, estramustine, fotemustine, glufosfamide, KW-2170,mafosfamide, mitolactol, lomustine, treosulfan, dacarbazine andtemozolomide.

Examples of antimetabolites include, but are not limited to,methotrexate, 6-mercaptopurine riboside, mercaptopurine, 5-fluorouracil(5-FU) alone or in combination with leucovorin, tegafur, UFT,doxifluridine, carmofur, cytarabine, cytarabine ocfosfate, enocitabine,S-1, pemetrexed, gemcitabine, fludarabine, 5-azacitidine, capecitabine,cladribine, clofarabine, decitabine, eflornithine, ethenylcytidine,cytosine arabinoside, hydroxyurea, TS-1, melphalan, nelarabine,nolatrexed, disodium pemetrexed, pentostatin, pelitrexol, raltitrexed,triapine, trimetrexate, vidarabine, mycophenolic acid, ocfosfate,pentostatin, tiazofurin, ribavirin, EICAR, hydroxyurea and deferoxamine.

Examples of antibiotics include, but are not limited to, intercalatingantibiotics, aclarubicin, actinomycin D, amrubicin, annamycin,adriamycin, bleomycin, daunorubicin, doxorubicin (including liposomaldoxorubicin), elsamitrucin, epirubicin, glarubicin, idarubicin,mitomycin C, nemorubicin, neocarzinostatin, peplomycin, pirarubicin,rebeccamycin, stimalamer, streptozocin, valrubicin, zinostatin andcombinations thereof.

Examples of topoisomerase inhibiting agents include, but are not limitedto, aclarubicin, amonafide, belotecan, camptothecin,10-hydroxycamptothecin, 9-amino-camptothecin, amsacrine, dexrazoxane,diflomotecan, irinotecan HCl, edotecarin, epirubicin, etoposide,exatecan, becatecarin, gimatecan, lurtotecan, orathecin, BN-80915,mitoxantrone, pirarbucin, pixantrone, rubitecan, sobuzoxane, SN-38,tafluposide and topotecan.

Examples of antibodies include, but are not limited to, rituximab,cetuximab, bevacizumab, trastuzumab, CD40-specific antibodies andIGF1R-specific antibodies, chTNT-1/B, denosumab, edrecolomab, WX G250,zanolimumab, lintuzumab and ticilimumab.

Examples of hormonal therapies include, but are not limited to,sevelamer carbonate, rilostane, luteinizing hormone releasing hormone,modrastane, exemestane, leuprolide acetate, buserelin, cetrorelix,deslorelin, histrelin, anastrozole, fosrelin, goserelin, degarelix,doxercalciferol, fadrozole, formestane, tamoxifen, arzoxifene,bicalutamide, abarelix, triptorelin, finasteride, fulvestrant,toremifene, raloxifene, trilostane, lasofoxifene, letrozole, flutamide,megesterol, mifepristone, nilutamide, dexamethasone, prednisone andother glucocorticoids.

Examples of retinoids or deltoids include, but are not limited to,seocalcitol, lexacalcitol, fenretinide, aliretinoin, tretinoin,bexarotene and LGD-1550.

Examples of plant alkaloids include, but are not limited to,vincristine, vinblastine, vindesine and vinorelbine.

Examples of proteasome inhibitors include, but are not limited to,bortezomib, MG-132, NPI-0052 and PR-171.

Examples of immunologicals include, but are not limited to, interferonsand numerous other immune-enhancing agents. Interferons includeinterferon alpha, interferon alpha-2a, interferon alpha-2b, interferonbeta, interferon gamma-1a, interferon gamma-1b, interferon gamma-n1 andcombinations thereof. Other agents include filgrastim, lentinan,sizofilan, BCG live, ubenimex, WF-10 (tetrachlorodecaoxide or TCDO),aldesleukin, alemtuzumab, BAM-002, dacarbazine, daclizumab, denileukin,gemtuzumab ozogamicin, ibritumomab, imiquimod, lenograstim, melanomavaccine, molgramostim, sargaramostim, tasonermin, tecleukin, thymalasin,tositumomab, Virulizin™ immunotherapeutic of Lorus Pharmaceuticals,Z-100 (specific substance of Maruyama or SSM), Zevalin™ (⁹⁰Y-ibritumomabtiuxetan), epratuzumab, mitumomab, oregovomab, pemtumomab, Provenge™(sipuleucel-T), teceleukin, Therocys™ (Bacillus Calmette-Guerin),cytotoxic lymphocyte antigen 4 (CTLA4) antibodies and agents capable ofblocking CTLA4 such as MDX-010.

Examples of biological response modifiers are agents that modify defensemechanisms of living organisms or biological responses, such assurvival, growth, or differentiation of tissue cells to direct them tohave anti-tumor activity. Such agents include, but are not limited to,krestin, lentinan, sizofuran, picibanil, PF-3512676 and ubenimex.

Examples of pyrimidine analogs include, but are not limited to,5-fluorouracil, floxuridine, doxifluridine, raltitrexed, cytarabine,cytosine arabinoside, fludarabine, triacetyluridine, troxacitabine andgemcitabine.

Examples of purine analogs include, but are not limited to,mercaptopurine and thioguanine.

Examples of antimitotic agents include, but are not limited to,N-(2-((4-hydroxyphenyl)amino)pyridin-3-yl)-4-methoxybenzenesulfonamide,paclitaxel, docetaxel, larotaxel, epothilone D, PNU-100940, batabulin,ixabepilone, patupilone, XRP-9881, vinflunine and ZK-EPO (syntheticepothilone).

Examples of radiotherapy include, but are not limited to, external beamradiotherapy (XBRT), teletherapy, brachytherapy, sealed-sourceradiotherapy and unsealed-source radiotherapy.

BiTE antibodies are bi-specific antibodies that direct T-cells to attackcancer cells by simultaneously binding the two cells. The T-cell thenattacks the target cancer cell. Examples of BiTE antibodies include, butare not limited to, adecatumumab (Micromet MT201), blinatumomab(Micromet MT103) and the like. Without being limited by theory, one ofthe mechanisms by which T-cells elicit apoptosis of the target cancercell is by exocytosis of cytolytic granule components, which includeperforin and granzyme B. In this regard, Bcl-2 has been shown toattenuate the induction of apoptosis by both perforin and granzyme B.These data suggest that inhibition of Bcl-2 could enhance the cytotoxiceffects elicited by T-cells when targeted to cancer cells (Sutton et al.(1997) J. Immunol. 158:5783-5790).

SiRNAs are molecules having endogenous RNA bases or chemically modifiednucleotides. The modifications do not abolish cellular activity, butrather impart increased stability and/or increased cellular potency.Examples of chemical modifications include phosphorothioate groups,2′-deoxynucleotide, 2′-OCH₃-containing ribonucleotides,2′-F-ribonucleotides, 2′-methoxyethyl ribonucleotides, combinationsthereof and the like. The siRNA can have varying lengths (e.g., 10-200bps) and structures (e.g., hairpins, single/double strands, bulges,nicks/gaps, mismatches) and are processed in cells to provide activegene silencing. A double-stranded siRNA (dsRNA) can have the same numberof nucleotides on each strand (blunt ends) or asymmetric ends(overhangs). The overhang of 1-2 nucleotides can be present on the senseand/or the antisense strand, as well as present on the 5′- and/or the3′-ends of a given strand. For example, siRNAs targeting Mcl-1 have beenshown to enhance the activity of ABT-263 (Tse et al. (2008), supra, andreferences therein).

Multivalent binding proteins are binding proteins comprising two or moreantigen binding sites. Multivalent binding proteins are engineered tohave the three or more antigen binding sites and are generally notnaturally occurring antibodies. The term “multispecific binding protein”means a binding protein capable of binding two or more related orunrelated targets. Dual variable domain (DVD) binding proteins aretetravalent or multivalent binding proteins binding proteins comprisingtwo or more antigen binding sites. Such DVDs may be monospecific (i.e.,capable of binding one antigen) or multispecific (i.e., capable ofbinding two or more antigens). DVD binding proteins comprising twoheavy-chain DVD polypeptides and two light-chain DVD polypeptides arereferred to as DVD Ig's. Each half of a DVD Ig comprises a heavy-chainDVD polypeptide, a light-chain DVD polypeptide, and two antigen bindingsites. Each binding site comprises a heavy-chain variable domain and alight-chain variable domain with a total of 6 CDRs involved in antigenbinding per antigen binding site.

PARP inhibitors include, but are not limited to, ABT-888, olaparib,KU-59436, AZD-2281, AG-014699, BSI-201, BGP-15, INO-1001, ONO-2231 andthe like.

Additionally or alternatively, a composition of the invention, forexample such a composition comprising ABT-263, can be administered incombination therapy with one or more antitumor agents selected fromABT-100, N-acetylcolchinol-O-phosphate, acitretin, AE-941, aglyconprotopanaxadiol, arglabin, arsenic trioxide, AS04 adjuvant-adsorbed HPVvaccine, L-asparaginase, atamestane, atrasentan, AVE-8062, bosentan,canfosfamide, Canvaxin™, catumaxomab, CeaVac™, celmoleukin, combrestatinA4P, contusugene ladenovec, Cotara™, cyproterone, deoxycoformycin,dexrazoxane, N,N-diethyl-2-(4-(phenylmethyl)phenoxy)ethanamine,5,6-dimethylxanthenone-4-acetic acid, docosahexaenoic acid/paclitaxel,discodermolide, efaproxiral, enzastaurin, epothilone B, ethynyluracil,exisulind, falimarev, Gastrimmune™, GMK vaccine, GVA™, halofuginone,histamine, hydroxycarbamide, ibandronic acid, ibritumomab tiuxetan,IL-13-PE38, inalimarev, interleukin 4, KSB-311, lanreotide,lenalidomide, lonafarnib, lovastatin, 5,10-methylenetetrahydrofolate,mifamurtide, miltefosine, motexafin, oblimersen, OncoVAX™ Osidem™,paclitaxel albumin-stabilized nanoparticle, paclitaxel poliglumex,pamidronate, panitumumab, peginterferon alia, pegaspargase, phenoxodiol,poly(I)-poly(C12U), procarbazine, ranpirnase, rebimastat, recombinantquadrivalent HPV vaccine, squalamine, staurosporine, STn-KLH vaccine, T4endonuclase V, tazarotene,6,6′,7,12-tetramethoxy-2,2′-dimethyl-1β-berbaman, thalidomide,TNFerade™, ¹³¹I-tositumomab, trabectedin, triazone, tumor necrosisfactor, Ukrain™, vaccinia-MUC-1 vaccine, L-valine-L-boroproline,Vitaxin™, vitespen, zoledronic acid and zorubicin.

In one embodiment, a composition of the invention, for example such acomposition comprising ABT-263, is administered in a therapeuticallyeffective amount to a subject in need thereof to treat a disease duringwhich is overexpressed one or more of antiapoptotic Bcl-2 protein,antiapoptotic Bcl-X_(L) protein and antiapoptotic Bcl-w protein.

In another embodiment, a composition of the invention, for example sucha composition comprising ABT-263, is administered in a therapeuticallyeffective amount to a subject in need thereof to treat a disease ofabnormal cell growth and/or dysregulated apoptosis.

Examples of such diseases include, but are not limited to, cancer,mesothelioma, bladder cancer, pancreatic cancer, skin cancer, cancer ofthe head or neck, cutaneous or intraocular melanoma, ovarian cancer,breast cancer, uterine cancer, carcinoma of the fallopian tubes,carcinoma of the endometrium, carcinoma of the cervix, carcinoma of thevagina, carcinoma of the vulva, bone cancer, colon cancer, rectalcancer, cancer of the anal region, stomach cancer, gastrointestinal(gastric, colorectal and/or duodenal) cancer, chronic lymphocyticleukemia, acute lymphocytic leukemia, esophageal cancer, cancer of thesmall intestine, cancer of the endocrine system, cancer of the thyroidgland, cancer of the parathyroid gland, cancer of the adrenal gland,sarcoma of soft tissue, cancer of the urethra, cancer of the penis,testicular cancer, hepatocellular (hepatic and/or biliary duct) cancer,primary or secondary central nervous system tumor, primary or secondarybrain tumor, Hodgkin's disease, chronic or acute leukemia, chronicmyeloid leukemia, lymphocytic lymphoma, lymphoblastic leukemia,follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin,melanoma, multiple myeloma, oral cancer, non-small-cell lung cancer,prostate cancer, small-cell lung cancer, cancer of the kidney and/orureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasmsof the central nervous system, primary central nervous system lymphoma,non Hodgkin's lymphoma, spinal axis tumors, brain stem glioma, pituitaryadenoma, adrenocortical cancer, gall bladder cancer, cancer of thespleen, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastomaor a combination thereof.

In a more particular embodiment, a composition of the invention, forexample such a composition comprising ABT-263, is administered in atherapeutically effective amount to a subject in need thereof to treatbladder cancer, brain cancer, breast cancer, bone marrow cancer,cervical cancer, chronic lymphocytic leukemia, acute lymphocyticleukemia, colorectal cancer, esophageal cancer, hepatocellular cancer,lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies ofT-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oralcancer, ovarian cancer, non-small-cell lung cancer, prostate cancer,small-cell lung cancer or spleen cancer.

According to any of these embodiments, the composition can beadministered in monotherapy or in combination therapy with one or moreadditional therapeutic agents.

For example, a method for treating mesothelioma, bladder cancer,pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous orintraocular melanoma, ovarian cancer, breast cancer, uterine cancer,carcinoma of the fallopian tubes, carcinoma of the endometrium,carcinoma of the cervix, carcinoma of the vagina, carcinoma of thevulva, bone cancer, colon cancer, rectal cancer, cancer of the analregion, stomach cancer, gastrointestinal (gastric, colorectal and/orduodenal) cancer, chronic lymphocytic leukemia, acute lymphocyticleukemia, esophageal cancer, cancer of the small intestine, cancer ofthe endocrine system, cancer of the thyroid gland, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, testicular cancer,hepatocellular (hepatic and/or biliary duct) cancer, primary orsecondary central nervous system tumor, primary or secondary braintumor, Hodgkin's disease, chronic or acute leukemia, chronic myeloidleukemia, lymphocytic lymphoma, lymphoblastic leukemia, follicularlymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma,multiple myeloma, oral cancer, non-small-cell lung cancer, prostatecancer, small-cell lung cancer, cancer of the kidney and/or ureter,renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of thecentral nervous system, primary central nervous system lymphoma, nonHodgkin's lymphoma, spinal axis tumors, brain stem glioma, pituitaryadenoma, adrenocortical cancer, gall bladder cancer, cancer of thespleen, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastomaor a combination thereof in a subject comprises administering to thesubject therapeutically effective amounts of (a) a composition of theinvention, for example such a composition comprising ABT-263, and (b)one or more of etoposide, vincristine, CHOP, rituximab, rapamycin,R-CHOP, RCVP, DA-EPOCH-R or bortezomib.

In particular embodiments, a composition of the invention, for examplesuch a composition comprising ABT-263, is administered in atherapeutically effective amount to a subject in need thereof inmonotherapy or in combination therapy with etoposide, vincristine, CHOP,rituximab, rapamycin, R-CHOP, RCVP, DA-EPOCH-R or bortezomib in atherapeutically effective amount, for treatment of a lymphoid malignancysuch as B-cell lymphoma or non-Hodgkin's lymphoma.

In other particular embodiments, a composition of the invention, forexample such a composition comprising ABT-263, is administered in atherapeutically effective amount to a subject in need thereof inmonotherapy or in combination therapy with etoposide, vincristine, CHOP,rituximab, rapamycin, R-CHOP, RCVP, DA-EPOCH-R or bortezomib in atherapeutically effective amount, for treatment of chronic lymphocyticleukemia or acute lymphocytic leukemia.

The present invention also provides a method for maintaining inbloodstream of a human cancer patient a therapeutically effective plasmaconcentration of ABT-263 and/or one or more metabolites thereof,comprising administering to the subject an ABT-263 composition asdescribed herein, in a dosage amount of about 50 to about 500 mg ABT-263free base equivalent per day, at an average dosage interval of about 3hours to about 7 days.

What constitutes a therapeutically effective plasma concentrationdepends inter alia on the particular cancer present in the patient, thestage, severity and aggressiveness of the cancer, and the outcome sought(e.g., stabilization, reduction in tumor growth, tumor shrinkage,reduced risk of metastasis, etc.). It is strongly preferred that, whilethe plasma concentration is sufficient to provide benefit in terms oftreating the cancer, it should not be sufficient to provoke an adverseside-effect to an unacceptable or intolerable degree.

Further information of relevance to the present invention is availablein a recently published article by Tse et al. (2008) Cancer Res.68:3421-3428 and supplementary data thereto available at Cancer ResearchOnline (cancerres.aacrjournals.org/). This article and its supplementarydata are incorporated in their entirety herein by reference.

EXAMPLES

The following examples are illustrative of the invention or of problemsovercome by the invention, but are not to be construed as limiting.Characterization of a particular embodiment as unfavorable or notselected for preparation of a prototype formulation does not necessarilymean that such embodiment is totally inoperative or outside the scope ofthe invention. One of skill in the art, based on the full disclosureherein, can prepare acceptable formulations even using ingredients shownherein to be suboptimal.

Trademarked ingredients used in the examples, which can be substitutedwith comparable ingredients from other suppliers, include:

-   -   Avicel 101™ and Avicel 102™ of FMC: microcrystalline cellulose;    -   Imwitor 742™ of Sasol: caprylic/capric mono- and diglycerides;    -   Miglyol 810™ of Sasol: caprylic/capric triglycerides;    -   Capmul MCM™ of Abitec: glyceryl caprylate/caprate;    -   Capmul PG-8™ of Abitec: propylene glycol monocaprylate;    -   Capmul PG12™ of Abitec: propylene glycol monolaurate;    -   Captex 300™ of Abitec: caprylic/capric triglycerides;    -   Cremophor EL™ of BASF: polyoxyethylene (35) castor oil;    -   Cremophor RH40™ of BASF: polyoxyethylene (40) hydrogenated        castor oil;    -   Crillet 4HP™ of Croda: polysorbate 80 having low peroxide value;    -   Gelucire 44/14™ of Gattefossé: polyoxyethylene glyceryl laurate;    -   Phosal 53 MCT™ of Phospholipid GmbH: blend containing not less        than 53% phosphatidylcholine, not more than 6%        lysophosphatidylcholine, about 29% medium chain triglycerides,        3-6% ethanol, about 3% mono- and diglycerides from sunflower        oil, about 2% oleic acid, and about 0.2% ascorbyl palmitate;    -   Plurol Oleique CC497™ of Gattefossé: polyglyceryl oleate;    -   ProSolv HD 90™ of JRS Pharma: silicified microcrystalline        cellulose;    -   Labrafil M 1944 CS™ of Gattefossé: polyoxyethylene glyceryl        monooleate;    -   Labrafil M 2125 CS™ of Gattefossé: polyoxyethylene glyceryl        linoleate;    -   Labrasol™ of Gattefossé: polyoxyethylene glyceryl        caprylate/caprate;    -   Lauroglycol 90™ of Gattefossé: propylene glycol monolaurate;    -   Lipoid S75™ MCT (prepared from Lipoid S75™ of Lipoid GmbH):        blend containing not less than 20% phosphatidylcholine, 2-4%        phosphatidylethanolamine, not more than 1.5%        lysophosphatidylcholine, and 67-73% medium-chain triglycerides;    -   Span™ 20 of Croda International PLC: sorbitan monolaurate;    -   Starch 1500™ of Colorcon: pregelatinized starch;    -   Tween™ 20 of Uniqema: polysorbate 20;    -   Tween™ 80 of Uniqema: polysorbate 80;    -   Vitamin E TPGS™: α-tocopheryl polyethylene glycol (1000)        succinate (TPGS).

All ABT-263 amounts, including concentrations and doses, given in theexamples are expressed as free base equivalent doses unless expresslystated otherwise. Where ABT-263 is administered as bis-HCl salt, 1.076mg ABT-263 bis-HCl provides 1 mg ABT-263 free base equivalent.

Example 1 Solubility of ABT-263 Parent and bis-HCl Salt in LipidSolvents

Solubility of ABT-263 parent (free base, crystalline Form I) and ABT-263bis-HCl salt was tested in a variety of lipid solvents and solventmixtures in ambient conditions. “PE-91” is Phosal 53 MCT™+ethanol, 9:1by volume. “LOT-343” is Labrafil M 1944 CS™+oleic acid+Tween 80™,30:40:30 by weight.

Solubility data are presented in Table 4. In some cases, indicated inTable 4 by an asterisk (*), solubility was initially high butprecipitation occurred upon standing.

TABLE 4 Solubility (mg/g) of ABT-263 parent and bis-HCl salt in lipidsolvents Solvent Parent (Form I) bis-HCl salt corn oil <86 <104 sesameoil <75 <80 castor oil * >78.8 Miglyol 810 ™ <76 <84 Lipoid S75 ™ MCT150-200 48.9 Phosal 53 MCT ™ >300 n.d. oleic acid >514 <498 Imwitor742 ™ * >245 Capmul MCM ™ * >321 Capmul PG-8 ™ * <43 Capmul PG-12 ™ *<39 Captex 300 ™ * <52 Labrafil M 1944 CS ™ >265 <45 Labrafil M 2125CS ™ >290 <44 PEG-400 >200 >278 propylene glycol * >337 Tween ™20 >256 >176 Tween ™ 80 >256 >125 Labrasol ™ >242 >292 CremophorRH40 ™ >226 n.d. poloxamer 124 >231 <41 PE-91 >250 89 LOT-343 >479 n.d.n.d. not determined

Example 2 Miscibility of Ternary Excipient Systems with ABT-263 Parentand bis-HCl Salt

Ternary systems consisting of two solvents and a surfactant wereevaluated for miscibility and drug solubility using 20% by weightABT-263 free base or 10% by weight ABT-263 bis-HCl salt. Solventsevaluated included Labrafil M 1944 CS™, Imwitor 742™ oleic acid, CapmulPG-8™, Capmul PG-12™, Lauroglycol 90™ and Phosal 53 MCT™. Surfactantsevaluated included Tween™ 80, Cremophor RH40™, Gelucire 44/14™ andLabrasol™. Data are presented in Table 5.

TABLE 5 Miscibility of ternary systems and solubility of ABT-263 parentand bis-HCl salt Miscibility ABT-263 solubility % by of 20% free Ternarysystem weight excipients 10% salt base Labrafil M 1944 CS ™ 30:45:25 ✓ ✓X Imwitor 742 ™ 40:35:25 ✓ ✓ X Tween 80 ™ 30:40:30 ✓ ✓ X (LIT systems)40:30:30 ✓ ✓ X Labrafil M 1944 CS ™ 30:45:25 ✓ ✓ ✓ oleic acid 40:35:25 ✓✓ ✓ Tween 80 ™ 30:40:30 ✓ ✓ ✓ (LOT systems) 40:30:30 ✓ ✓ ✓ Capmul PG-8 ™45:30:25 ✓ X X Labrafil M 1944 CS ™ 35:40:25 ✓ X X Tween 80 ™ 40:30:30 ✓X X (C8LT systems) 30:40:30 ✓ X X Capmul PG-12 ™ 45:30:25 ✓ ✓ ✓ LabrafilM 1944 CS ™ 35:40:25 ✓ ✓ ✓ Tween 80 ™ 40:30:30 ✓ ✓ ✓ (C12LT systems)30:40:30 ✓ ✓ ✓ Imwitor 742 ™ 45:30:25 X N/A (vehicle not miscible)Labrafil M 1944 CS ™ 35:40:25 X N/A (vehicle not miscible) CremophorRH40 ™ 40:30:30 X N/A (vehicle not miscible) (ILC systems) 30:40:30 XN/A (vehicle not miscible) 60:30:10 ✓ ✓ X 50:40:10 ✓ ✓ X 50:30:20 ✓ ✓ X40:40:20 ✓ ✓ X Labrafil M 1944 CS ™ 30:45:25 X N/A (vehicle notmiscible) oleic acid 40:35:25 X N/A (vehicle not miscible) CremophorRH40 ™ 30:40:30 X N/A (vehicle not miscible) (LOC systems) 40:30:30 XN/A (vehicle not miscible) 30:60:10 ✓ ✓ ✓ 40:50:10 ✓ ✓ ✓ 30:50:20 X N/A(vehicle not miscible) 40:40:20 X N/A (vehicle not miscible) CapmulPG-8 ™ 45:30:25 X N/A (vehicle not miscible) Labrafil M 1944 CS ™35:40:25 X N/A (vehicle not miscible) Cremophor RH40 ™ 40:30:30 X N/A(vehicle not miscible) (C8LC systems) 30:40:30 X N/A (vehicle notmiscible) 60:30:10 ✓ X X 50:40:10 ✓ X X 50:30:20 ✓ X X 40:40:20 ✓ X XCapmul PG-12 ™ 45:30:25 X N/A (vehicle not miscible) Labrafil M 1944CS ™ 35:40:25 X N/A (vehicle not miscible) Cremophor RH40 ™ 40:30:30 XN/A (vehicle not miscible) (C12LC systems) 30:40:30 X N/A (vehicle notmiscible) Lauroglycol 90 ™ 45:30:25 ✓ ✓ ✓ Labrafil M 1944 CS ™ 35:40:25X N/A (vehicle not miscible) Cremophor RH40 ™ 40:30:30 X N/A (vehiclenot miscible) (LLC systems) 30:40:30 X N/A (vehicle not miscible)Imwitor 742 ™ 60:30:10 X N/A (vehicle not miscible) Labrafil M 1944 CS ™50:40:10 X N/A (vehicle not miscible) Gelucire 44/14 ™ 50:30:20 X N/A(vehicle not miscible) (ILG systems) 40:40:20 X N/A (vehicle notmiscible) oleic acid 60:30:10 X N/A (vehicle not miscible) Labrafil M1944 CS ™ 50:40:10 X N/A (vehicle not miscible) Gelucire 44/14 ™50:30:20 X N/A (vehicle not miscible) (OLG systems) 40:40:20 X N/A(vehicle not miscible) Capmul PG-8 ™ 60:30:10 X N/A (vehicle notmiscible) Labrafil M 1944 CS ™ 50:40:10 X N/A (vehicle not miscible)Gelucire 44/14 50:30:20 X N/A (vehicle not miscible) (C8LG systems)40:40:20 X N/A (vehicle not miscible) Lauroglycol 90 ™ 60:30:10 X N/A(vehicle not miscible) Labrafil M 1944 CS ™ 50:40:10 X N/A (vehicle notmiscible) Gelucire 44/14 ™ 50:30:20 X N/A (vehicle not miscible) (LLGsystems) 40:40:20 X N/A (vehicle not miscible) Imwitor 742 ™ 60:30:10 ✓✓ X Labrafil M 1944 CS ™ 50:40:10 ✓ ✓ X Labrasol ™ 50:30:20 ✓ ✓ X (ILLsystems) 40:40:20 ✓ ✓ X oleic acid 60:30:10 ✓ ✓ ✓ Labrafil M 1944 CS ™50:40:10 ✓ ✓ ✓ Labrasol ™ 50:30:20 ✓ ✓ ✓ (OLL systems) 40:40:20 ✓ ✓ ✓Capmul PG-8 60:30:10 ✓ X X Labrafil M 1944 CS ™ 50:40:10 ✓ X XLabrasol ™ 50:30:20 ✓ X X (C8LL systems) 40:40:20 ✓ ✓ ✓ Lauroglycol 90 ™60:30:10 ✓ ✓ X Labrafil M 1944 CS ™ 50:40:10 ✓ ✓ X Labrasol ™ 50:30:20 ✓✓ ✓ (LLL systems) 40:40:20 ✓ ✓ ✓

All ternary excipient systems tested containing 10-20% Gelucire 44/14™exhibited immiscibility. Most systems tested containing greater than 20%Cremophor RH40™ also showed immiscibility. Only in certain systems wherethe excipients were miscible was ABT-263 in free base or bis-HCl saltform soluble at the concentrations tested.

Data for further ternary systems containing phosphatidylcholine-basedexcipients are presented in Example 8, Tables 11 and 12.

Example 3 Chemical Stability of ABT-263 Free Base and bis-HCl Salt inLipid Solution

Preliminary stability studies were conducted to allow a side-by-sidecomparison between lipid solutions of ABT-263 in bis-HCl salt and freebase form. ABT-263 was dissolved in two separate sets of lipid vehicles,Phosal 53 MCT™/ethanol (9:1 by volume; “PE-91”) and Labrafil M 1944CS™/oleic acid/Tween 80™ (30:40:30 by weight; “LOT-343”). No antioxidantwas included, nor was headspace nitrogen purging performed. After agingof samples at 40° C. (stress condition) for up to 3 weeks, analysis oftotal sulfoxides indicated that free base was significantly more stablethan bis-HCl salt in the solutions tested (Table 6). Total degradantlevels also showed a similar trend (data not shown). The increase indegradant level was accompanied by color change. The bis-HCl saltsolutions upon aging showed pronounced color darkening whereas the freebase solutions exhibited very little color change.

TABLE 6 Sulfoxide formation in lipid solutions of ABT-263 free base andbis-HCl salt % w/w total sulfoxides Solution A Solution B Time free basebis-HCl salt free base bis-HCl salt (weeks) 25 mg/ml 25 mg/ml 100 mg/ml100 mg/ml 0 0.05 0.07 2.49 2.24 1 0.27 0.79 3.70 7.15 2 0.53 1.90 4.1137.52  3 0.84 3.44 no data no data

Example 4 Chemical Stability of ABT-263 Free Base in Various LipidSolutions

The chemical stability of the ABT-263 free base in solution in variouslipid excipients was assessed by conducting a two-week stress test at40° C., without antioxidant or nitrogen purging. Results are presentedin Table 7.

TABLE 7 Sulfoxide formation in lipid solutions of ABT-263 free base %w/w total Concentration sulfoxides* Lipid solvent (mg/g) Initial 1 week2 weeks Lipoid S75 ™ MCT 100 0.21 0.33 0.51 Imwitor 742 ™   25** 0.250.20 0.14 Capmul PG-8 ™   25** 0.21 0.25 0.19 Tween 80 ™ 100 0.20 0.590.84 Crillet 4HP ™ 100 0.18 0.44 0.64 Plurol Oleique   50** 0.31 2.416.26 CC497 ™/Lipoid S75 ™ MCT 50:50 v/v Labrafil M 1944 CS ™ 100 0.305.86 9.16 oleic acid (super-refined) 100 0.04 0.18 0.29 Phosal 53 MCT ™/ 50 n.d. 0.14 0.18 ethanol 9:1 v/v *sulfoxide was analyzed as peak %relative to that of ABT-263 **lower concentration was used due to lowdrug solubility in the lipid vehicle n.d. not detectable

The following can be summarized from the above study.

-   -   Very little or only slight growth of sulfoxides was seen in        phosphatidylcholine-based lipid excipients such as Phosal 53        MCT™ or Lipoid S75™ MCT.    -   Very little or only slight growth of sulfoxides was seen in        Imwitor 742™, Capmul PG-8™ and oleic acid (super-refined grade).    -   Moderate sulfoxide growth was seen in Tween 80™. The degradation        was slowed down when a purer grade of polysorbate 80 (Crillet        4HP™) was used.    -   Labrafil M 1944 CS™ and Plurol Oleique CC497™ were both        associated with significant degradation of the ABT-263. Both        these excipients contain oleic acid in their structure, and the        unsaturated nature of oleic acid is known to promote oxidative        reaction. This may be the reason for the chemical instability of        the drug in these excipients.

Example 5 Chemical Stability of ABT-263 Free Base in Ternary LipidSolution Systems

Although ABT-263 appeared to be stable in super-refined oleic acidduring the two-week stressed test of Example 4, a subsequent test usingmulticomponent vehicles showed that drug solutions containing oleic acidled to color change upon standing. A comparative storage study wasconducted at ambient temperature using solutions of ABT-263 in Imwitor742™/oleic acid/Tween 80™ (30:40:30 by weight; “IOT-343”) and Imwitor742™/Phosal 53 MCT™/Tween 80™ (40:40:20 by weight; “IPT-442”). TheIOT-343 vehicle itself was colorless, and adding ABT-263 free base at10% by weight to the vehicle only made it very slightly yellow-hued, butthe color of the resulting ABT-263 solution darkened significantly uponstorage. This was in contrast to a solution of ABT-263 free base at 10%by weight in IPT-442 solution, which had a yellow colored vehicle tobegin with, but only darkened slightly upon storage. HPLC analysis forthe two drug solutions after storage at ambient conditions for 3 monthsconfirmed that the color change correlated to degradation (totalsulfoxide levels were 1.3% for the IOT-343 system and 0.5% for theIPT-442 system). Therefore, oleic acid was excluded from lipidexcipients to be used for ABT-263 liquid-filled capsule formulation.

Further stress testing on ABT-263 free base lipid solutions usingdifferent ternary lipid combinations showed that Labrafil M 1944 CS™ wasalso associated with significant oxidative degradation of ABT-263. Asshown by results from a three-week stress test presented in Table 8,formulations containing Labrafil M 1944 CS™ showed significant sulfoxidegrowth upon storage at 40° C. without antioxidant or nitrogen purging.On the other hand, an Imwitor 742™/Phosal 53 MCT™/Tween 80™ (20:50:30 byweight; “IPT-253”) solution of ABT-263 which had neither oleic acid norLabrafil M 1944 CS™ showed much enhanced chemical stability compared tothe other formulations tested, namely Labrafil M 1944 CS™/oleicacid/Tween 80™ (30:40:30 by weight; “LOT-343”) and Labrafil M 1944CST™/Imwitor 742™/Tween 80™ (40:30:30 by weight; “LIT-433”). Therefore,both Labrafil M 1944 CS™ as well as oleic acid was excluded from lipidexcipients to be used for ABT-263 liquid-filled capsule formulation.

TABLE 8 Sulfoxide formation in ternary lipid solutions of ABT-263 freebase Ternary lipid Concentration % w/w total sulfoxides* solvent system(mg/g) Initial 1 week 2 weeks 3 weeks LOT-343 100 2.49 3.70 4.11 no dataLIT-433 100 0.21 3.20 5.13 no data LIT-433 150 0.23 2.28 3.61 3.80IPT-253 150 n.d. 0.26 0.47 0.56 *sulfoxide was analyzed as peak %relative to that of ABT-263 n.d. not detectable

Example 6 Antioxidant Testing for ABT-263 Free Base in Lipid SolutionSystems

The effectiveness of different antioxidants in inhibiting oxidativedegradation was evaluated in lipid solutions containing ABT-263 freebase at 100 mg/g in two different lipid solution systems: (1) LipoidS75™ MCT and (2) a ternary lipid system (LIT-433; see above). The latterwas purposely chosen as a system promoting significant degradation in ashort time, as an antioxidant screen. Sulfoxide formation during thetwo-week stress test at 40° C. with nitrogen purging is shown in Table9.

TABLE 9 Effect of antioxidants on sulfoxide formation in solutions ofABT-263 free base % w/w total sulfoxides* Antioxidant In Lipoid S75 ™MCT In LIT-433 Antioxidant concentration Initial 1 week 2 weeks Initial1 week 2 weeks none 0.06 0.42 0.68 0.21 3.20 5.13 ascorbyl palmitate100% molar** n.d. n.d. n.d. 0.31 1.37 2.07 BHA 100% molar** 0.13 0.260.30 0.43 2.25 3.66 BHT 100% molar** 0.08 0.17 0.27 0.37 2.07 3.40 Nametabisulfite***  0.1% (w/w) cloudy solution 0.18 1.95 3.07 Nathiosulfate***  0.1% (w/w) cloudy solution 0.18 2.64 4.31 thioglycerol100% molar** 0.08 0.09 0.13 0.33 0.50 0.56 α-tocopherols 145% molar**0.20 0.27 0.50 0.41 3.99 9.23 n.d. not determined (ascorbyl palmitatecould not be dissolved at 100% relative molar concentration in thissolvent) *sulfoxide was analyzed as peak % relative to that of ABT-263**molar concentration relative to ABT-263 ***an aqueous stock solutionof 15% w/v was prepared for antioxidant addition.

ABT-263 free base degraded to a much lesser extent in the Lipoid S75™MCT vehicle than in the LIT-433 vehicle system. Thioglycerol providedeffective inhibition of drug oxidation in both vehicle systems. In theLIT-433 vehicle system, ascorbyl palmitate, butylated hydroxyanisole(BHA), butylated hydroxytoluene (BHT), sodium metabisulfite and sodiumthiosulfate inhibited oxidative degradation to some extent at theconcentrations tested, but α-tocopherols were ineffective. It is notedthat the concentrations of sodium metabisulfite and sodium thiosulfatewere very much lower than those providing molar equivalence to ABT-263.Even at the low concentrations used, the addition of water with theseantioxidants led to cloudy solutions. The concentrations of ascorbylpalmitate, BHA and BHT were much higher than typically used forantioxidant purposes.

Example 7 BHA as an Antioxidant for ABT-263 Free Base in Ternary LipidSolution Systems

Due to its favorable lipophilic nature and wide use in lipid system asan antioxidant, the antioxidant effectiveness of BHA was tested, at aconcentration more typical for BHA, in two additional ternary vehiclesystems, IPT-253 and LIT-433, containing ABT-263 at 150 mg/g. Testingwas done in stress conditions at 40° C. without nitrogen purging. Asshown in Table 10, in both systems, addition of 0.2% w/w BHA did notprovide any inhibition of sulfoxide formation. It was concluded thatfree-radical-scavenger types of antioxidant such as BHA and BHT do notappear to be useful in protecting ABT-263 from oxidative degradation inlipid solutions.

TABLE 10 Effect of BHA on sulfoxide formation in solutions of ABT-263free base Ternary % w/w total sulfoxides system Antioxidant Initial 1week 2 weeks 3 weeks 4 weeks IPT-253 none n.d. 0.26 0.47 0.56 0.67 0.2%w/w BHA 0.06 0.29 0.49 0.58 0.68 LIT-433 none 0.23 2.28 3.61 3.86 4.190.2% w/w BHA 0.24 2.22 3.54 3.80 4.19 n.d. not detectable

Example 8 Phospholipid Solution Systems for ABT-263 Free Base

Based on the above studies, the phosphatidylcholine-containingexcipients Phosal 53 MCT™ and Lipoid S75™ MCT were concluded to providegood chemical stability and drug solubility for ABT-263 free base.However, these pre-blended excipients are not suitable for use alone asa vehicle for an ABT-263 liquid-filled capsule, due to either highviscosity (Phosal 53 MCT™) or insufficient drug solubility (Lipoid S75™MCT). Polysorbate 80 could be used to enhance drug solubility in thevehicle. Excipients such as Capmul PG8™ or Imwitor 742™ could be used toreduce viscosity of the lipid solution. Both were shown to be chemicallycompatible with ABT-263. Imwitor 742™ was preferred over Capmul PG8™based on previous experience in FDA approved drug products.

Consequently, in developing a prototype liquid-filled capsule, attentionfocused on excipients such as Phosal 53 MCT™, Lipoid S75™ MCT,polysorbate 80 (the purer forms such as Crillet 4HP™ and super-refinedTween 80™ being preferred) and Imwitor 742™

Two ternary lipid vehicle systems containing either Imwitor 742™/Phosal53 MCT™/Tween 80™ (abbreviated as “IPT”) systems or Imwitor 742™/LipoidS75™ MCT/Tween 80™ (abbreviated as “IST”) systems at various excipientratios were investigated in a screen for prototype capsule formulations.The level of Imwitor 742™ in the ternary blend was limited to no morethan 40%, and the level of polysorbate 80 to no more than 20%. Thethree-digit suffix following “IPT” or “IST” refers to the respectivepercentages of the three excipient ingredients, in each case omittingthe final zero.

Selection of prototype formulations was based on vehicle miscibility,ABT-263 free base solubility in the vehicle, viscosity of the resultingsolution (judged by severity of stringing when released from a dropper)and self-dispersing property of the drug solution (at 10% by weight drugloading), as summarized in Tables 11 and 12 for IPT and IST systemsrespectively. Schematic phase diagrams for IPT and IST systems (FIGS. 1and 2) further illustrate the selection process.

As can be seen from Tables 11 and 12 and the phase diagrams in FIGS. 1and 2, the IPT systems in general provided better vehicle miscibility,drug solubility and dispersibility than the corresponding IST systems.IPT-262 and IST-262 (later replaced by IST-172) were selected asprototype vehicle systems, based on the following rationales.

A phosphatidylcholine-based solvent (for example in the form of Phosal53 MCT™ or Lipoid S75™ MCT) is needed to ensure both chemical stability(and bioavailability—see below) of the capsule formulation. The amountof such solvent is virtually unlimited due to the low toxicity and hightolerance of lecithin used in oral products.

Polysorbate 80 (especially grades of high purity) is needed tofacilitate drug solubility in the vehicle and to enhanceself-dispersibility of the lipid formulation. Based on a typical dailydose of ABT-263 (e.g., 200-250 mg) and a maximum daily dose ofpolysorbate 80 (418 mg), it is reasonable to limit the level ofpolysorbate 80 to no more than 20% in the vehicle for a prototypeformulation with 10% drug loading. Higher levels of polysorbate 80 arealso unfavorable due to chemical stability considerations.

In the IPT systems, Imwitor 742™ is needed to reduce the viscosity ofthe final drug solution to a level that allows for machine capsulefilling. In the IST system, Imwitor 742™ is also needed to enhance themiscibility of the vehicle system, since Lipoid S75™ MCT and polysorbate80 are not miscible at all ratios. However, the amount of Imwitor 742™is limited to no more than 20% in both prototype systems.

It will be noted from Table 12 that the IST-172 system exhibits poorvehicle miscibility. However, it was found that upon addition of ABT-263free base the miscibility of the entire system was acceptable; thus theIST-172 formulation became an acceptable prototype liquid forencapsulation.

TABLE 11 Formulation properties of IPT systems containing 10% ABT-263free base Vehicle Drug Vehicle miscibility solubility Stringing*Dispersibility (description) IPT-190 ✓ ✓ ++ Dispersed with vigorousshaking IPT-280 ✓ ✓ ++ Dispersed with vigorous shaking IPT-370 ✓ ✓ ++Dispersed with gentle shaking IPT-460 ✓ ✓ + Dispersed with gentleshaking IPT-091 ✓ ✓ +++ Dispersed with vigorous shaking IPT-181 ✓ ✓ ++Dispersed with vigorous shaking IPT-271 ✓ ✓ + Dispersed with vigorousshaking IPT-361 ✓ ✓ + Dispersed with vigorous shaking IPT-451 ✓ ✓ −Dispersed with gentle shaking IPT-082 ✓ ✓ +++ Dispersed with vigorousshaking IPT-172 ✓ ✓ ++ Dispersed with gentle shaking IPT-262 ✓ ✓ +Dispersed with gentle shaking IPT-352 ✓ ✓ + Dispersed with gentleshaking IPT-442 ✓ ✓ − Dispersed with gentle shaking ✓ vehicle miscible,or drug fully dissolved in vehicle *stringing: +++ extreme; ++significant; + slight; − none

TABLE 12 Formulation properties of IST systems containing 10% ABT-263free base Vehicle Drug Vehicle miscibility solubility Stringing*Dispersibility (description) IST-190 ✓ ✓ − Oil drops spread but did notdisperse until shaken vigorously IST-280 ✓ ✓ − Oil drops spread but didnot disperse until shaken vigorously IST-370 ✓ X n/a n/a IST-460 ✓ X n/an/a IST-091 X ✓ n/a n/a IST-181 X ✓ − Dispersed with gentle shakingIST-271 ✓ ✓ − Dispersed with gentle shaking IST-361 ✓ X n/a n/a IST-451✓ X n/a n/a IST-082 X n/a n/a n/a IST-172 X ✓ ++ Rapidly dispersed withgentle shaking IST-262 ✓ ✓ + Rapidly dispersed with gentle shakingIST-352 ✓ ✓ + Dispersed with gentle shaking IST-442 ✓ X n/a n/a ✓vehicle miscible, or drug fully dissolved in vehicle X vehicleimmiscible or miscible but turbid, or residual solids present (due toundissolved drug or precipitation) n/a solution not made due toimmiscible vehicle, or dispersibility test not performed due toundissolved drug *stringing: +++ extreme; ++ significant; + slight; −none

Example 9 Antioxidant Selection for Phospholipid-Based Solutions ofABT-263 Free Base

Based on initial antioxidant screening (see Example 6), acceleratedstability studies were further conducted on the two prototypeformulations using either sodium metabisulfite (NaMTBS) or thioglycerolas an antioxidant, together with 0.01% EDTA.

The solubility of neat NaMTBS in IPT-262 and IST-262 solutionscontaining 10% ABT-263 free base and 0.01% EDTA (as edetate calciumdisodium) was assessed. After 5 days of rotary mixing under ambienttemperature conditions, solids remained in all solutions, at NaMTBSsolid concentrations as low as 0.05% w/w (or approximately 2% molarconcentration relative to ABT-263).

Due to poor lipid solubility of NaMTBS, an alternative way ofintroducing it to the lipid solution is by adding a concentrated aqueousstock solution of NaMTBS to the lipid solution. For example, a clearsolution was obtained when a 50 mg/ml free base solution in Phosal 53MCT™/ethanol 9:1 v/v was spiked with a 15% w/v NaMTBS solution up to afinal NaMTBS concentration of 9.67 mg/ml (or 100% molar concentrationrelative to ABT-263). However, as the final concentration of NaMTBS wasincreased to 150% relative molar concentration or higher, using the 15%w/v stock solution, the lipid solution turned turbid. Using a stocksolution at a concentration greater than 20% also results in solutionturbidity, indicating that both excess amounts of water and NaMTBS canlead to a cloudy solution.

Example 10 Sulfoxide Formation in Phospholipid-Based FormulationsContaining Antioxidant

Results from a two-week accelerated stability study (stress condition:40° C., with nitrogen purging), as shown in Table 13, indicated thatthioglycerol is not as effective as NaMTBS in inhibiting sulfoxideformation in both prototype formulations.

However, the study results also showed that water added with the NaMTBScan negatively impact chemical stability of the drug solution, and thishas been shown to be the case regardless of the ABT-263 form (free baseor bis-HCl salt) or the vehicle system used (see Table 14; two-weekstudy at 40° C., with nitrogen purging). For this reason, a finalconcentration of 0.05% (w/w) NaMTBS was selected, and the concentrationof MTBS stock solution should also be kept below about 15% w/v in orderto avoid turbidity.

TABLE 13 Sulfoxide formation in ABT-263 prototype liquids forencapsulation % water % w/w total sulfoxides Vehicle Antioxidant added*initial 1 week 2 weeks IST-172 none 0 0.06 0.34 0.54 IST-172 0.05%NaMTBS + 0.32 0.19 0.28 0.22 0.01% EDTA IST-172 0.55% Thioglycerol + 00.22 0.27 0.55 0.01% EDTA IPT-262 none 0 0.14 0.41 0.55 IPT-262 0.05%NaMTBS + 0.32 0.43 0.31 0.23 0.01% EDTA IPT-262 0.55% Thioglycerol + 00.11 0.26 0.42 0.01% EDTA *water as % of formulation contributed by 15%w/v NaMTBS stock solution

TABLE 14 Sulfoxide formation in ABT-263 lipid solutions: effects ofNaMTBS and water ABT-263 % w/w total Vehicle ABT-263 form concentrationantioxidant water % sulfoxides PE-91 free base 50 mg/ml none 0 0.47(Form I) PE-91 free base 50 mg/ml none 3.00 0.66 (Form I) PE-91 bis-HCl50 mg/ml none 0 1.90 salt PE-91 bis-HCl 50 mg/ml 0.05% NaMTBS + 0.320.53 salt 0.01% EDTA PE-91 bis-HCl 50 mg/ml  0.1% NaMTBS + 0.61 0.84salt 0.01% EDTA PE-91 bis-HCl 50 mg/ml  0.2% NaMTBS + 1.17 0.97 salt0.01% EDTA IST-172 free base 100 mg/g none 0 0.54 (Form I) IST-172 freebase 100 mg/g 0.05% NaMTBS + 0.32 0.22 (Form I) 0.01% EDTA IST-172 freebase 100 mg/g  0.1% NaMTBS + 0.61 0.22 (Form I) 0.01% EDTA IST-172 freebase 100 mg/g  0.2% NaMTBS + 1.17 0.58 (Form I) 0.01% EDTA IPT-262 freebase 100 mg/g none 0 0.55 (Form I) IPT-262 free base 100 mg/g 0.05%NaMTBS + 0.32 0.23 (Form I) 0.01% EDTA IPT-262 free base 100 mg/g  0.1%NaMTBS + 0.61 0.37 (Form I) 0.01% EDTA IPT-262 free base 100 mg/g  0.2%NaMTBS + 1.17 0.58 (Form I) 0.01% EDTA

Example 11 In Vivo Pharmacokinetics of Prototype Liquid-Filled Capsules

Two 100 mg/g ABT-263 free base liquid-filled capsule prototypeformulations were dosed in dogs (single-dose, non-fasting conditions) toevaluate their in vivo pharmacokinetics in comparison with 50 mg/ml oralsolutions of ABT-263 free base and bis-HCl salt in Phosal 53MCT™/ethanol 9:1 v/v with 0.01% EDTA. Formulations tested were:

-   -   Formulation 3: 100 mg/g ABT-263 free base in Imwitor 742™/Phosal        53 MCT™/Tween 80™ 20:60:20 (“IPT-262”), liquid-filled capsule;    -   Formulation 4: 100 mg/g ABT-263 free base in Imwitor 742™/Lipoid        S75™ MCT/Tween 80™ 20:60:20 (“IST-262”), liquid-filled capsule;    -   Formulation 5: 50 mg/ml ABT-263 free base in Phosal 53        MCT™/ethanol 9:1 v/v, oral solution; and    -   Formulation 6: 50 mg/ml ABT-263 bis-HCl in Phosal 53        MCT™/ethanol 9:1 v/v, oral solution.

Each formulation was evaluated in a group of six dogs at a dose of 50mg/dog. Formulations 3 (IPT-262) and 4 (IST-262) were dosed in the samegroup of dogs in a sequential manner, and Formulations 5 and 6 weredosed in a separate group of dogs in a sequential manner. The dogs werefasted overnight prior to dosing, but food was provided 30 minutes priorto dosing. Plasma concentrations of parent drug were determined byHPLC-MS/MS at the completion of each study. Results are presented inTable 15.

TABLE 15 Dog pharmacokinetics of prototype liquid-filled capsules (3 and4) versus comparative liquid formulations (5 and 6) C_(max) T_(max) AUCFormulation (μg/ml) (h) (μg · h/ml) F % 3 9.8 4.7 98.6 41.9 4 11.0 2.576.8 31.8 5 11.3 6.0 107.8 42.5 6 11.9 4.5 94.1 37.7

The peak concentration (C_(max)) of Formulation 3 in plasma was slightlylower than that of Formulation 4, but AUC of Formulation 3 was higherthan that of Formulation 4, apparently due to slower absorption.Formulation 4 showed a more consistent but shorter T_(max) of 2-3 hoursafter dosing. Liquid-filled capsule Formulation 3 gave comparable plasmaC_(max), AUC and bioavailability (F %) to that of the oral solutions(Formulations 5 and 6). Based on these results, the IPT-262 prototype(Formulation 3) was selected as a liquid-filled capsule formulation forhuman clinical studies.

Example 12 Storage Stability of Prototype Formulations with and withoutNaMTBS

Preliminary physical and chemical stability results have been obtainedon two laboratory-scale batches of a prototype ABT-263 liquid-filledcapsule formulation. The only difference between the two batches ispresence or absence of antioxidant (sodium metabisulfite). Compositionof the two batches is shown in Table 16.

TABLE 16 Composition of prototype liquid for capsules used in stabilitystudy Batch 1 Batch 2 (with antioxidant) (without antioxidant) mg per mgper Component capsule % w/w capsule % w/w ABT-263 free base 50.0 10.050.0 10.0 sodium metabisulfite 0.25 0.05 — — edetate calcium disodium0.025 0.005 0.025 0.005 water* 2.48 0.50 0.23 0.05 Phosal 53 MCT ™268.35 53.67 269.85 53.97 mono- and dicaprylic/capric 89.45 17.89 89.9517.99 glycerides polysorbate 80 89.45 17.89 89.95 17.99 Total 500.0100.0 500.0 100.0 *includes water added with sodium metabisulfite andedetate calcium disodium only

The liquids having the composition shown in Table 16 were encapsulatedin size 0 hard gelatin capsules and the capsules placed in blisterpackaging for a chemical stability study. Data after one month storageunder various conditions are presented in Table 17. Water content shownin Table 17 is as determined by analysis, and is not directly related toamount of water added with NaMTBS and edetate calcium disodium as inTable 16.

It can be seen from Table 17 that addition of the antioxidant sodiummetabisulfite significantly inhibited formation of total sulfoxides,especially under stress storage conditions of 40° C. and 75% RH.

TABLE 17 Chemical stability results for prototype capsules with andwithout antioxidant initial 1 month water water Storage total totalcontent total total content Batch conditions sulfoxides degradants (%)*sulfoxides degradants (%) 1 (with  5° C. n.d. 0.03% 2.7 n.d. 0.03% 3.1antioxidant) 25° C. n.d. 0.03% 2.7 n.d. 0.06% 3.6 60% RH 40° C. n.d.0.03% 2.7 n.d. 0.03% 4.8 75% RH 2 (without  5° C. 0.08% 0.14% 3.2 0.12%0.17% 3.3 antioxidant) 25° C. 0.08% 0.14% 3.2 0.08% 0.11% 3.1 60% RH 40°C. 0.08% 0.14% 3.2 0.29% 0.42% 3.8 75% RH *Initial water content of fillsolution: 0.4% for batch 1; 0.2% for batch 2 n.d. not detectable

Example 13 Preparation of an Illustrative Nanoparticulate Suspension

ABT-263 nanoparticulate suspension formulations were prepared byhigh-pressure homogenization as described below. The formulations hadthe following compositions (all percentages expressed as weight/volume)in water:

Formulation 7 ABT-263 bis-HCl   5% (4.65% free base equivalent)poloxamer 188   3% Formulation 8 ABT-263 bis-HCl   5% (4.65% free baseequivalent) poloxamer 188   3% NaHCO₃ 8.4%

Aqueous solutions were prepared containing the indicated amount ofpoloxamer 188 (Pluronic™ F68) and, in the case of Formulation 8, sodiumbicarbonate (NaHCO₃). Crystalline ABT-263 bis-HCl in an amountsufficient to provide a 5% weight/volume (50 mg/ml) suspension wasdispersed in each aqueous solution using a Sonifier™ homogenizer(Branson Ultrasonic, Danbury, Conn.). The resulting dispersion was thenadded to the sample reservoir of a Microfluidizer™ M-110L processor(Microfluidics International Corp., Newton, Mass.) and processed at12,000 psi (approximately 82.5 MPa) for 2 hours. The sample temperaturewas maintained throughout at a temperature of 20±2° C. by running thedispersion through a heat exchanger immersed in a water bath connectedto a chiller.

The suspensions so obtained (Formulations 7 and 8) were subjected toparticle size measurement immediately upon preparation and after storagefor 14 days at 5° C. (see Example 14). Formulation 8 was submitted to anoral pharmacokinetic (PK) study in dogs (see Example 15).

Example 14 Effect of Sodium Bicarbonate on Particle Size Stability ofNanosuspensions

Formulations 7 and 8 were compared as to their particle sizedistribution (D₉₀ and D₅₀). Particle size measurement was performedimmediately upon preparation of the suspensions (t=0) and after storagefor 14 days at 5° C. In addition particle size was measured at t=0 forsuspensions following dilution of 1 ml of each suspension in 20 ml 0.9%sodium chloride (NaCl) solution. Data are given in Table 18.

TABLE 18 D₉₀ and D₅₀ particle sizes (μm) of nanosuspension Formulations7 and 8 Formulation 7 Formulation 8 (no NaHCO₃) (8.4% NaHCO₃) D₉₀ D₅₀D₉₀ D₅₀ t = 0 1.126 0.490 0.605 0.291 14 d at 5° C. 1.214 0.570 0.6210.295 t = 0 in 0.9% NaCl 1.712 0.886 0.596 0.295

Example 15 Pharmacokinetics of an Illustrative Nanosuspension

Single-dose pharmacokinetics of Formulation 8 of Example 13 wereevaluated in non-fasted beagle dogs (n=4) after a 5 mg/kg oral dose. Theformulation was administered in two ways: by oral gavage and in acapsule. Formulation 8 was also administered to histamine-pretreatedfasted dogs (n=4), by oral gavage only. For comparative purposes, asolution formulation of ABT-263 bis-HCl in a lipid medium (FormulationC, prepared from ABT-263 bis-HCl powder dissolved to a concentration of25 mg/ml in a 90:10 mixture of Phosal 53 MCT™ and ethanol) wasadministered to non-fasted dogs. Formulation C has been used to evaluateABT-263 in clinical studies.

Serial heparinized blood samples were obtained from a jugular vein ofeach animal prior to dosing and 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 9, 12, 15and 24 hours after administration. Plasma was separated bycentrifugation (2,000 rpm for 10 minutes at approximately 4° C.) andABT-263 was isolated using protein precipitation with acetonitrile.

ABT-263 and an internal standard were separated from each other and fromco-extracted contaminants on a 50×3 mm Keystone Betasil CN™ 5 μm columnwith an acetonitrile/0.1% trifluoroacetic acid mobile phase (50:50 byvolume) at a flow rate of 0.7 ml/min. Analysis was performed on a SciexAPI3000™ biomolecular mass analyzer with a heated nebulizer interface.ABT-263 and internal standard peak areas were determined using SciexMacQuan™ software. The plasma drug concentration of each sample wascalculated by least squares linear regression analysis (non-weighted) ofthe peak area ratio (parent/internal standard) of the spiked plasmastandards versus concentration. The plasma concentration data weresubmitted to multi-exponential curve fitting using WinNonlin 3(Pharsight).

The area under the plasma concentration-time curve from 0 to t hours(time of the last measured plasma concentration, which here is 24 hours)after dosing (AUC₀₋₂₄) was calculated using the linear trapezoidal rulefor the plasma concentration-time profiles.

Mean plasma concentrations over 24 hours after dosing are shown in FIG.3.

Calculated mean PK parameters are summarized in Table 19.

TABLE 19 PK parameters (mean ± SEM) in dogs (non-fasted unless otherwiseindicated) C_(max) T_(max) AUC₀₋₂₄ Bioavailability (μg/ml) (h) (μg ·h/ml) F % Formulation C 9.09 ± 1.33 6.3 ± 1.6 54.5 ± 6.3 22.4 ± 2.6(comparative) Formulation 8, 7.78 ± 0.35 2.3 ± 0.3 45.2 ± 2.6 19.9 ± 1.2oral gavage Formulation 8, 7.52 ± 2.46 3.0 ± 0.4  48.3 ± 12.4 21.3 ± 5.5in capsule Formulation 8, 5.56 ± 0.46 3.3 ± 0.3 35.6 ± 0.6 15.7 ± 0.2oral gavage (fasted dogs)

Example 16 Preparation of Solid Dispersions of ABT-263 bis-Hcl

ABT-263 bis-HCl crystalline salt was mixed with a surfactant and awater-soluble polymer in the following weight ratios:

10.8% ABT-263 salt (10% free base equivalent); 10% surfactant; 79.2%polymer

21.5% ABT-263 salt (20% free base equivalent); 10% surfactant; 68.5%polymer

32.3% ABT-263 salt (30% free base equivalent); 10% surfactant; 57.7%polymer

43% ABT-263 salt (40% free base equivalent); 10% surfactant; 47% polymer

The surfactant in different series was TPGS, Span™ 20 or Tween™ 20. Thepolymer in different series was copovidone (Kollidon™ VA 64), povidoneK-30 or HPMC-AS.

The mixture of ingredients in each case was dissolved in methanol. Themethanol was removed at 65° C. in vacuo using a Genevac™ system, and theresulting solid dispersion was allowed to cool to ambient temperature.

The solid dispersion in each case was sieved through a 40-mesh screen toprovide a powder of reduced particle size. The resulting powders wereused for determination of T_(g) by differential scanning calorimetry(DSC), residual solvent and moisture determination by thermogravimetricanalysis (TGA), characterization of crystallinity or lack thereof bypowder X-ray diffraction (PXRD), and determination of physical stabilitywhen stored at 25° C./60% RH and at 40° C./75% RH.

The solid dispersion powder in each case was blended with ProSolv HD90™, croscarmellose sodium and sodium stearyl fumarate at a weight ratioof 82:15:2:1. The resulting blend was filled into hard gelatin capsulesof a size, depending on drug loading, to provide a 50 mg unit dose ofABT-263. The capsules were tested for dissolution in a pH 6.5 buffermedium containing 7.6 mM Tween™ 80, using USP apparatus II (see Example17 below).

All tested solid dispersions of ABT-263 bis-HCl prepared as above werefound to have a T_(g) in the range of 70-110° C. TGA showed that thecopovidone/HPMC-AS dispersions had the lowest moisture content (2-4%)and the povidone dispersions, regardless of surfactant used, had thehighest moisture content (8-10%). PXRD showed no crystallinity, i.e.,the ABT-263 bis-HCl was amorphous in all solid dispersions. Only theABT-263 bis-HCl solid dispersions prepared with HPMC-AS as the polymericcarrier showed acceptable storage stability for one month. Wherepovidone or copovidone was used, a tendency for deliquescence wasobserved in open-dish storage stability testing at both at 25° C./60% RHand at 40° C./75% RH.

Example 17 Preparation of Solid Dispersions of ABT-263 Free Base

ABT-263 bis-HCl crystalline salt was dissolved in acetone, and NaOH wasadded to convert the ABT-263 bis-HCl to free base. The NaCl by-productprecipitated and was removed by filtration.

To the resulting ABT-263 free base solution in acetone were added asurfactant and a water-soluble polymer in the following weight ratios:

10% ABT-263 free base; 10% surfactant; 80% polymer

20% ABT-263 free base; 10% surfactant; 70% polymer

30% ABT-263 free base; 10% surfactant; 60% polymer

40% ABT-263 free base; 10% surfactant; 50% polymer

The surfactant in different series was TPGS, Span™ 20 or Tween™ 20. Thepolymer in different series was copovidone (Kollidon™ VA 64) or HPMC-AS.

The acetone was removed at 65° C. in vacuo using a Genevac™ system, andthe resulting solid dispersion was allowed to cool to ambienttemperature.

The solid dispersion in each case was sieved through a 40-mesh screen toprovide a powder of reduced particle size. The resulting powders, as inExample 16, were used for determination of T_(g) by DSC, residualsolvent and moisture determination by TGA, characterization ofcrystallinity or lack thereof by PXRD, and determination of physicalstability when stored at 25° C./60% RH and at 40° C./75% RH.

The solid dispersion powder in each case was blended with ProSolv HD90™, croscarmellose sodium and sodium stearyl fumarate at a weight ratioof 82:15:2:1. The resulting blend was filled into hard gelatin capsulesof a size, depending on drug loading, to provide a 50 mg unit dose ofABT-263. The capsules were tested for dissolution in a pH 6.5 buffermedium containing 7.6 mM Tween™ 80 (see Example 18 below).

All tested solid dispersions of ABT-263 free base prepared as above werefound to have a T_(g) in the range of 70-110° C. TGA showed that thecopovidone and HPMC-AS dispersions had low moisture content (2-4%). PXRDshowed no crystallinity, i.e., the ABT-263 free base was amorphous inall solid dispersions. The ABT-263 free base solid dispersions preparedwith copovidone or HPMC-AS as the polymeric carrier showed acceptablestorage stability for one month without any sign of deliquescence.

Example 18 Dissolution Profiles of Solid Dispersions

Representative dissolution (drug release) profiles in a pH 6.5 bufferedmedium containing 7.6 mM Tween™ 80 are shown in FIG. 4 (ABT-263 bis-HCl)and FIG. 5 (ABT-263 free base).

As shown in FIG. 4, at a 20% drug-loading level, the ABT-263 bis-HClsolid dispersions with 68.5% copovidone and 10% TPGS showed a moderaterate of drug release that plateaued at about 80% release. Release fromsimilar dispersions having Span™ 20 or, especially, Tween™ 20 as thesurfactant was much slower.

By contrast, as shown in FIG. 5, at the same 20% drug-loading level, theABT-263 free base solid dispersions with 70% copovidone and 10% ofeither Tween™ 20 or TPGS showed rapid dug release. Only the Span™ 20surfactant resulted in much slower release in the case of the free basedispersion.

Release rate was drug-loading-dependent in both ABT-263 bis-HCl and freebase dispersion formulations, the 20% dispersions showing faster releasethan the 30% or 40% dispersions in both cases.

Unlike the analogous solid dispersion prepared from the ABT-263 freebase, the solid dispersion containing ABT-263 bis-HCl, copovidone andTween™ 20 showed shell formation. This shell formation is believed to becaused by precipitation of the drug on the surface of the capsule fillplug.

In a separate study, solid dispersions of ABT-263 bis-HCl in acopovidone matrix with and without replacement of 5% copovidone withHPMC-AS showed slower drug release in presence of HPMC-AS.

Example 19 Effect of Polymeric Carrier on Dissolution Profile of ABT-263bis-HCl Dispersions

Solid dispersions with different polymeric carriers were tested toobserve impact of the polymeric carriers on dissolution rates. Foursolid dispersions were prepared with ABT-263 bis-HCl salt (20% free baseequivalent), 10% TPGS and the following polymeric carriers:

povidone only

50% povidone+50% copovidone

25% povidone+75% copovidone

copovidone only

Dissolution profiles of the four solid dispersions are shown in FIG. 6.Drug release rate increased with increasing levels of povidone.

Example 20 Pharmacokinetics of ABT-263 bis-HCl Dispersions in a DogModel

Single-dose pharmacokinetics of two ABT-263 solid dispersions wereevaluated in non-fasted beagle dogs (n=6) after a 50 mg/kg oral dosefollowed by 10 ml water. Serial heparinized blood samples were obtainedfrom a jugular vein of each animal prior to dosing and 0.25, 0.5, 1,1.5, 2, 3, 4, 6, 9, 12, 15 and 24 hours after administration. Plasma wasseparated by centrifugation (2,000 rpm for 10 minutes at approximately4° C.) and ABT-263 was isolated using protein precipitation withacetonitrile.

Two ABT-263 bis-HCl solid dispersions (those of Example 19 containingpovidone only or copovidone only) were compared. The powdereddispersions were blended with ProSolv HD 90™, croscarmellose sodium andsodium stearyl fumarate in an 82:15:2:1 weight ratio and the blendfilled into capsules.

ABT-263 and an internal standard were separated from each other and fromco-extracted contaminants on a 50×3 mm Keystone Betasil CN™ 5 μm columnwith an acetonitrile/0.1% trifluoroacetic acid mobile phase (50:50 byvolume) at a flow rate of 0.7 ml/min. Analysis was performed on a SciexAPI3000™ biomolecular mass analyzer with a heated nebulizer interface.ABT-263 and internal standard peak areas were determined using SciexMacQuan™ software. The plasma drug concentration of each sample wascalculated by least squares linear regression analysis (non-weighted) ofthe peak area ratio (parent/internal standard) of the spiked plasmastandards versus concentration. The plasma concentration data weresubmitted to multi-exponential curve fitting using WinNonlin 3(Pharsight).

The area under the plasma concentration-time curve from 0 to t hours(time of the last measured plasma concentration) after dosing(AUC_(0-t)) was calculated using the linear trapezoidal rule for theplasma concentration-time profiles. The residual area extrapolated toinfinity, determined as the final measured plasma concentration (C_(t))divided by the terminal elimination rate constant (β), was added toAUC_(0-t) to produce the total area under the curve (AUC_(0-∞)). Thebioavailability was calculated as the dose-normalized AUC_(0-∞)from oraldosing divided by the corresponding value derived from i.v.(intravenous) dosing, administered as a slow bolus to a jugular veinunder light ether anesthetic.

PK parameters for the povidone-only and copovidone-only dispersions arepresented in Table 20.

TABLE 20 PK parameters of solid dispersion compositions in dog (n = 6)C_(max)/D AUC/D C_(max) μg/ml per AUC μg · h/ml Composition μg/ml mg/kgT_(max) h μg · h/ml per mg/kg F % povidone 5.6 1.16 9.8 39.3 7.9 16.4copovidone 9.6 1.78 4.5 64.9 11.9 24.7

Although the ABT-263 bis-HCl dispersion prepared with povidone was shownin Example 19 to provide a better release rate than copovidone, it hadpoorer bioavailability in this dog study than a comparable dispersionprepared with copovidone.

Example 21 Pharmacokinetics of Illustrative Solid Dispersions in a DogModel

Single-dose pharmacokinetics of two ABT-263 solid dispersions wereevaluated in non-fasted beagle dogs (n=6), following the same protocolas that of Example 20. Two ABT-263 solid dispersions (Dispersions I andII) were prepared. Dispersion I, prepared substantially according to theprocess of Example 17, contained 10% ABT-263 free base, 10% TPGS and 80%copovidone. The powdered dispersion was filled into capsules without anyadditional ingredients to prepare Formulation 9. Dispersion II, preparedsubstantially according to the process of Example 16, contained 13.11%ABT-263 bis-HCl (12.18% free base equivalent), 15% TPGS and 71.89%povidone. The powdered dispersion was blended with ProSolv HD 90™,sodium starch glycolate and sodium stearyl fumarate in an 82:15:2:1weight ratio and the blend filled into capsules to prepare Formulation10.

PK parameters for Formulations 9 and 10 are presented in Table 21.

TABLE 21 PK parameters of solid dispersion compositions in dog (n = 6)C_(max)/D AUC/D C_(max) μg/ml per AUC μg · h/ml Formulation μg/ml mg/kgT_(max) h μg · h/ml per mg/kg F % 9 7.5 1.50 8.5 59.0 11.2 24.6 10 6.41.24 7.8 39.2 7.4 16.3

The ABT-263 bis-HCl dispersion (Formulation 10) prepared with povidonehad poorer bioavailability in this dog study than the ABT-263 free basedispersion (Formulation 9) prepared with copovidone.

Example 22 Preparation and Characterization of Solid Dispersion Products

Formulations of various compositions were produced as shown in Table 22below. ABT-263 was mixed in a blender with a pre-granulated mixture ofCopovidone (copolymer of N-vinyl pyrrolidone and vinyl acetate) and thesolubilizer(s). Where indicated, 1% of colloidal silicon dioxide wasadded to improve flow properties. The powdery mixture was extruded in aLeistritz micro 18 GMP-extruder at an extrusion temperature as shown inTable 22.

Absolute bioavailability compares the bioavailability (estimated as thearea under the curve, or AUC) of the active drug in systemic circulationfollowing oral administration with the bioavailability of the same drugfollowing intravenous administration. In Table 22 the bioavailability (F%) was determined after administering an ABT-263 dose of 50 mg to feddogs.

TABLE 22 Composition, stability and biovailability in dogs of soliddispersions Formulation 11 12 13 14 15 16 17 18 ABT-263 bis-HCl (%) 1010.7 10.7 10.7 10 10 10 10 copovidone (%) 80 72.3 72.3 72.3 80 79 80 79polysorbate 20 (%) 10 10 5 Span ™ 20 (%) 5 5 Vitamin E-TPGS ™ (%) 10 2 55 sodium lauryl sulfate (%) 6 6 6 propylene glycol (%) 3 5 5 5 5colloidal silicon dioxide (%) 1 1 1 1 1 extrusion temperature (° C.) 140140 140 140 140 140 130 130 sum of degradation products (%) 1.83 1.111.22 1.05 2.78 1.07 1.68 0.93 sum of sulfoxides (%) n.d. 0.77 0.71 0.69n.d. n.d. n.d. n.d. bioavailability (F %) 27.5 32.6 25.9 27.0 31.7 n.d.26.7 n.d. F % for Formulation C 22.4 31.5 29.2 29.2 22.4 n.d. 22.4 n.d.in same study relative F %** 122.8 103.5 88.7 92.5 141.5 n.d. 119.2 n.d.Formulation 19 20 21 22 23 24 25 ABT-263 form and amount (%) bis- bis-bis- Na free free bis- HCl HCl HCl salt base base HCl 10.7 10.7 10.7 1010 10 10.7 copovidone (%) 78.3 78.3 72.3 79 79 79 72.3 polysorbate 20(%) 10 Span ™ 20 (%) 10 Vitamin E-TPGS ™ (%) 5 5 5 10 10 sodium laurylsulfate (%) 5 6 propylene glycol (%) 5 5 5 colloidal silicon dioxide (%)1 1 1 1 1 1 1 extrusion temperature (° C.) 130 135 140 130 125 130 130sum of degradation products (%) 0.66 0.83 1.23 0.73 0.80 0.41 1.27 sumof sulfoxides (%) 0.37 0.42 0.72 0.29 0.43 0.30 0.62 bioavailability (F%) n.d. 29.6 n.d. 32.1 33.7 n.d. n.d. F % for Formulation C in samestudy n.d. n.d. n.d. 31.5 n.d. n.d. n.d. relative F %** n.d. n.d. n.d.101.9 n.d. n.d. n.d. n.d. not determined **calculated by takingbioavailability (F %) for Formulation C as 100%

Example 23 Bioavailability Evaluation of Solid Dispersions (a) Protocolfor Oral Bioavailability Studies

For bioavailability evaluation, extrudates as described in Example 22were milled and filled into capsules. Each capsule contained 50 mgABT-263.

The dose response and food effect for two formulations were evaluated inbeagle dogs (both genders, approximate weight: 10 kg). Groups of 5 dogseach received a 50 mg (1 capsule/dog), 100 mg (2 capsules/dog) or 200 mg(4 capsules/dog) oral dose of ABT-263 under both fasting and fedconditions. The dose was followed by approximately 10 ml water. For allstudies, beagle dogs were fasted overnight prior to dosing, but werepermitted water ad libitum. Food was returned to the dogs approximately30 minutes prior to dosing (fed conditions) or 4 hours after dosing(fasting conditions). A washout/recovery period of one week separatedthe two dosing periods. Blood samples were obtained from each animalprior to dosing and at convenient time points chosen among 0.25, 0.5,1.0, 1.5, 2, 3, 4, 6, 9, 12, 15, 24, 36 and 48 hours after drugadministration. The plasma was separated from the red cells bycentrifugation and frozen at −30° C. until analysis. Concentrations ofABT-263 were determined by reverse phase HPLC-MS/MS followingliquid-liquid extraction of the plasma samples. The area under the curve(AUC) was calculated by the trapezoidal method over the time course ofthe study. Each dosage form was evaluated in a group containing 5 dogs;the values reported are averages for each group of dogs.

(b) Influence of Dosage and Application to Fasted or Fed Dogs

Formulations 16 or 18 of ABT-263 as defined in Table 22 wereadministered to fasted or fed dogs in dosages corresponding to theamounts of ABT-263 as indicated in FIG. 7 and FIG. 8. Subsequently, theplasma concentrations of ABT-263 were determined from blood samplestaken at the indicated time points. In FIG. 7 and FIG. 8, open andclosed symbols represent fed or fasted dogs, respectively. Squares,triangles and circles represent a dose of 50 mg, 100 mg or 200 mgABT-263, respectively.

For both formulations plasma concentrations of ABT-263 were higher whenadministered to fed dogs. This effect was more prominent at higherdosages of 100 mg and 200 mg. In fed dogs a dose linearity could beobserved. AUC values of Formulation 16 in fasted dogs were 40-60% lowerthan in fed dogs. When Formulation 18 was administered AUC values wereapproximately 30% lower in fasted dogs.

(c) Comparison of a Free Base Formulation Vs. a bis-HCl Salt Formulation

Fed dogs received orally one of the following two formulations as onecapsule containing Formulation 23 or Formulation 20 as indicated inTable 22, equivalent to an amount of 50 mg ABT-263.

The plasma concentrations of ABT-263 were determined from blood samplestaken at the time points as indicated in FIG. 9, which shows the meanplasma concentration of five dogs treated with Formulation 23 orFormulation 20, respectively.

The bioavailability data obtained from this experiment are summarized inTable 23 below (shown as mean value of 6 animals; standard deviation inbrackets).

TABLE 23 Pharmacokinetics in fed dogs of solid dispersion formulationsFormulation C_(max) C_(max)/D T_(max) AUC AUC/D F % 23 (free base) 10.4(2.1) 2.03 3.2 (0.5) 78.7 (15.7) 15.3 33.7 (5.5) 20 (bis-HCl salt)  8.6(0.7) 1.74 3.6 (0.6) 67.6 (7.9)  13.4 29.6 (3.4) C_(max) maximumconcentration of ABT-263 in plasma (μg/ml) C_(max)/D maximumconcentration per dose (μg/ml per mg/kg) T_(max) time to maximum plasmaconcentration (h) AUC area under the plasma concentration curve (μg ·hr/ml) AUC/D area under curve per dose (μg · hr/ml per mg/kg) F %average bioavailability

Example 24 Storage Stability

For selected formulations (Formulations 16 and 18 according to Table 22)the storage stability was determined. The formulations were kept inclosed containers at ambient conditions (approximately 19° C. to 25° C.at RH of 60% or less). The ABT-263 content and the content ofdegradation products of the active ingredient including sulfoxides weredetermined at the beginning of the storage period (initial value) andafter 4 months by separation via HPLC (or HPLC) and detection with aUV/VIS detector. The results are shown in Table 24 below.

TABLE 24 Storage stability of solid dispersion formulations Formulation16 18 ABT-263 content (initial) 97.8% 97.0% degradation products(initial) 1.07% 0.93% ABT-263 content (after 4 months) 96.7% 98.9%degradation products (after 4 months) 1.16% 0.96%

The formulations were chemically stable as content and impurity levelsremained unchanged upon storage.

Example 25 Determination of Sulfoxide Formation

Formulations 12, 13, 22, 14, 19, 21, 20, 23 and 24 as defined in Table22 were assessed for sulfoxide formation in an accelerated stabilitystudy, using exposure in an open dish at a relative humidity of 40°C./75%. Sulfoxide content was determined at the beginning of theexperiment (less than 0.8% in all cases), after 1 week, 3 weeks and 6weeks for the formulations referred to in FIG. 10, and at time pointschosen among 4 weeks, 5 weeks and 7 weeks for the formulations referredto in FIG. 11.

The data shown in FIG. 10 indicate that lower extrusion temperaturescause lower contents of sulfoxides. Comparatively low levels ofsulfoxides were also observed in the formulations referred to in FIG.11, all of which were extruded at temperatures of 135° C. or less.Sulfoxide contents increased most significantly with Formulations 12 and14, both of which contain polysorbate 20. Therefore, the inclusion ofpolysorbate 20 appears to promote formation of sulfoxides.

In a second experiment sulfoxide formation was determined in sampleswhich were kept in closed 1.5 oz HDPE bottles at a temperature andrelative humidity of 40° C./75%. The results are shown in FIG. 12 andFIG. 13.

Example 26 Crystallinity of ABT-263 Extrudates

Formulations 19, 12, 23 and 24 as defined in Table 22 were manufactured,using the process parameters as indicated in Table 25 below. Theextrudates were evaluated for the presence of crystalline activeingredient by polarization microscopy.

TABLE 25 Crystallinity of ABT-263 extrudates Formulation 20 23 24 25Process parameters: feed rate 0.5 kg/h 1.0 kg/h 1.0 kg/h 0.5 kg/htemperature 135° C. 125° C. 130° C. 130° C. Process data: crystallinitydetected not detected not detected not detected

Example 27 Crystallinity of ABT-263 Extrudates Upon Prolonged Storage

Various extrudates as indicated in Table 26 were kept at acceleratedaging conditions in open dishes or closed bottles. At the indicated timepoints the presence of crystalline active ingredient was evaluated bypolarization microscopy.

TABLE 26 Physical stability (crystallinity) of ABT-263 extrudates Time 0weeks 1 week 3 weeks 6 weeks 1 month Storage open dish 40° C./75% RH 1.5oz HDPE conditions bottles, closed, 40° C./75% RH 12 detected (++)detected (++) detected (++) detected (++) detected (++) 13 detected (++)detected (++) detected (++) detected (++) detected (++) 22 not detectednot detected not detected not detected not detected 14 not detecteddetected (+) detected (++) detected (++) detected (++) 19 detected (+)not detected not detected detected (+) detected (+) 21 detected (+)detected (++) detected (++) detected (++) detected (++) 20 detected (+)detected (+) detected (+) detected (+) detected (+) 23 not detected notdetected not detected not detected not detected 24 not detected notdetected not detected not detected not detected (+) few crystalsdetected (++) numerous crystals detected

Example 28 Manufacture of Tablets

Following the procedure of Example 22, an extrudate was obtained fromthe solid dispersion product ingredients listed in Table 27 below.Extrudates from Example 22 were milled and the powder was blended withthe tableting excipients listed in Table 27. A single-punch tablet presswas used to prepare tablets containing 50 mg ABT-263.

TABLE 27 Tablet composition Formulation 26 27 28 extrudate (ABT-263 free 98%  83%  83% base:copovidone:Vitamin E-TPGS ™:colloidal silicondioxide 10:79:10:1) croscarmellose sodium  15% mannitol  15% colloidalsilicon dioxide 1.0% 1.0% 1.0% sodium stearyl fumarate 1.0% 1.0% 1.0%Total tablet mass 510.2 mg 602.4 mg 602.4 mg

The tablets were immersed in 0.1N HCl at a temperature of 37° C. (tomimic stomach conditions) and stirred by paddle rotation at a speed of75 rpm. The amount of released ABT-263 was determined at various timepoints by HPLC-UV/VIS. The results are shown in FIG. 14.

Example 29 PK Studies of ABT-263 Solid Tablets in Dogs

PK studies were performed in non-fasting beagle dogs (n=3) at a singledose of 50 mg ABT-263 free base equivalent. Plasma concentrations of thedrug were determined by high pressure liquid chromatography massspectrometry (HPLC-MS) and PK parameters were calculated by standardprocedures in the art.

Eleven tablet compositions of the invention (Formulations 26-36) weretested. API (ABT-263 bis-HCl in all cases) was unmilled unless otherwiseindicated. Composition of each of Formulations 26-30 is as shown inTable 28.

TABLE 28 Composition of tablets (Formulations 26-30) Amount (% byweight) Ingredient 26 27 28 29 30 ABT-263 bis-HCl 10.00 10.00 10.0010.75 10.75 Avicel 101 ™ 81.25 84.25 50.75 30.00 30.00 mannitol 20.0040.00 40.00 povidone K-30 3.00 3.00 5.00 5.00 3.00 crospovidone 1.501.50 poloxamer (Pluronic ™ F127) 4.00 1.00 4.00 TPGS 4.00 6.00 sodiumstarch glycolate 10.00 10.00 10.00 magnesium stearate 0.25 0.25 0.250.25 0.25

Formulations 31-36 comprised intra- and extragranular components.Composition of each of these formulations is as shown in Table 29.

TABLE 29 Composition of tablets (Formulations 31-36) Amount (% byweight) Ingredient 31 32 33 34 35 36 Intragranular ABT-263 bis-HCl 10.7510.75 10.75 21.50 10.75 21.50 Avicel 101 ™ 33.00 34.00 30.00 29.25 30.0029.25 mannitol 20.00 20.00 20.00 20.00 30.00 20.00 PVP 30 5.00 5.00 5.005.00 5.00 5.00 poloxamer (Pluronic ™ F127) 1.00 sodium starch glycolate5.00 5.00 5.00 5.00 Cremophor EL ™ 4.00 4.00 TPGS 4.00 4.00Extragranular Avicel 101 ™ 20.00 20.00 20.00 10.00 20.00 20.00 sodiumstarch glycolate 5.00 5.00 5.00 5.00 5.00 5.00 magnesium stearate 0.250.25 0.25 0.25 0.25 0.25

Formulation 37 consists of the following ingredients (all percentages byweight):

ABT-263 bis-HCl 10.75% ProSolv HD 90 ™ 49.00% mannitol 20.00% Starch1500 ™ 5.00% sodium starch glycolate 10.00% poloxamer (Pluronic ™ F127)4.00% colloidal silicon dioxide 1.00% sodium stearyl fumarate 0.25%

Tablets were prepared by one of the processes shown in Table 30.

TABLE 30 Processes used in preparing tablets Process Description I Wetgranulation; API suspended in binder solution (PVP + poloxamer) II Wetgranulation; API blended intragranularly III Dry blend; directlycompressed tablets

Table 31 summarizes PK data for ABT-263 tablet formulations in dogs. F %is a measure of bioavailability.

TABLE 31 PK data for tablet formulations AUC Formulation Process T_(max)(h) C_(max) (μg/ml) (μg · h/ml) F % 26 I 5.3 ± 1.2 2.2 ± 1.0 24.1 9.6 I2.3 ± 0.6 3.5 ± 0.3 28.5 12.0 API jet-milled 27 I 7.0 ± 6.9 1.8 ± 0.520.1 8.3 II 3.0 ± 0.0 4.0 ± 1.1 37.7 16.8 28 I 7.3 ± 6.7 3.6 ± 1.6 47.721.5 29 II 6.7 ± 5.0 3.9 ± 2.2 37.5 14.9 30 II 1.8 ± 0.3 7.5 ± 2.3 60.722.6 31 II 2.7 ± 0.6 6.1 ± 2.5 47.6 20.6 32 II 2.3 ± 0.6 7.1 ± 3.2 42.618.6 33 II 4.3 ± 4.0 3.6 ± 1.1 34.5 13.6 34 II 3.7 ± 2.1 5.8 ± 1.5 48.319.2 35 II 3.0 ± 1.0 6.8 ± 1.3 69.9 25.5 36 II 3.0 ± 1.0 4.5 ± 3.2 51.720.4 37 III 3.0 ± 1.0 10.2 ± 2.9  76.2 31.0

Tablets prepared by direct compression (Process III) exhibited higherbioavailability in these dog studies than those prepared by wetgranulation (Processes I and II). Tablets prepared by Process IIgenerally provided higher bioavailability in dogs than those prepared byProcess I. Adding the drug by suspending it in the binder solution alsoappeared to prolong the T_(max).

Addition of a surfactant to tablets made by wet granulation did notsignificantly change in vivo absorption of the drug. Addition ofwater-soluble excipients such as mannitol appeared to enhance in vivodrug absorption.

A change in drug loading level (21.5% vs. 10.75% ABT-263 bis-HCl; 20%vs. 10% free base equivalent) did not significantly changebioavailability.

Increasing the binder (e.g., PVP) concentration for wet granulation hada tendency to reduce bioavailability.

Example 30 PK Studies of ABT-263 Solid Capsules in Dogs

PK studies were performed in non-fasting beagle dogs (n=3) at a singledose of 50 mg ABT-263 free base equivalent. Plasma concentrations of thedrug were determined by high pressure liquid chromatography massspectrometry (HPLC-MS) and PK parameters were calculated by standardprocedures in the art.

Four capsule compositions of the invention (containing Formulations38-41) were tested. API (ABT-263 bis-HCl in all cases) was unmilledunless otherwise indicated.

Formulation 38 consists of the following ingredients (all percentages byweight):

ABT-263 bis-HCl 10.75% ProSolv HD 90 ™ 49.00% mannitol 20.00% starch1500 5.00% sodium starch glycolate 10.00% poloxamer (Pluronic ™ F127)4.00% colloidal silicon dioxide 1.00% magnesium stearate 0.25%

Formulation 39 consists of an intragranular component and anextragranular component having the following ingredients (allpercentages by weight):

Intragranular ABT-263 bis-HCl 10.75% Avicel 101 ™ 30.00% mannitol 30.00%poloxamer (Pluronic ™ F127)  1.00% hydroxypropylcellulose  3.00% sodiumstarch glycolate  2.5% Extragranular Avicel 101 ™ 20.00% sodium starchglycolate  2.5% sodium stearyl fumarate  0.25%

Formulation 40 consists of the following ingredients (all percentages byweight):

ABT-263 bis-HCl 10.75% ProSolv HD 90 ™ 50.00% mannitol 30.00%hydroxypropylcellulose 3.00% poloxamer (Pluronic ™ F127) 1.00% sodiumstarch glycolate 5.00% sodium stearyl fumarate 0.25%

Formulation 41 consists of the following ingredients (all percentages byweight):

ABT-263 bis-HCl 16.12% Avicel 102 ™ 50.00% mannitol 28.13% sodium starchglycolate 5.00% colloidal silicon dioxide 0.50% sodium stearyl fumarate0.25%

Capsule fills were prepared by one of the processes shown in Table 32.

TABLE 32 Processes used in preparing capsules Process Description II Wetgranulation; API blended intragranularly IV Dry blend encapsulation

Table 33 summarizes PK data for ABT-263 tablet formulations in dogs.Formulation 41 was tested three times.

TABLE 33 PK data for capsule formulations C_(max) AUC FormulationProcess T_(max) (h) (μg/ml) (μg · h/ml) F % 38 IV 4.2 ± 2.4 6.3 ± 1.554.1 21.7 39 II 6.7 ± 5.4 4.7 ± 2.4 51.0 20.3 II 3.8 ± 1.3  45 ± 1.940.5 13.2 API jet-milled 40 III 3.2 ± 0.8 6.2 ± 2.0 53.0 21.0 III 4.7 ±3.7 7.4 ± 2.0 74.5 34.2 API jet-milled 41 IV 2.8 ± 0.7 2.5 ± 0.5 43.215.8 7.0 ± 4.8 5.0 ± 1.2 62.3 23.5 4.2 ± 1.5 6.4 ± 2.9 52.6 17.6

Micronization of the API by jet-milling led to improved bioavailabilityfor capsules made by dry blending (Process IV) but not by wetgranulation (Process II). Addition of poloxamer surfactant did notsignificantly affect bioavailability of a dry blend encapsulationformulation.

1. An orally deliverable pharmaceutical composition comprising as a soleor first active ingredient a compound of Formula I

where X³ is chloro or fluoro; and (1) X⁴ is azepan-1-yl, morpholin-4-yl,1,4-oxazepan-4-yl, pyrrolidin-1-yl, —N(CH₃)₂, —N(CH₃)(CH(CH₃)₂),7-azabicyclo[2.2.1]heptan-7-yl or 2-oxa-5-azabicyclo[2.2.1]hept-5-yl;and R⁰ is

where X⁵ is —CH₂—, —C(CH₃)₂— or —CH₂CH₂—; X⁶ and X⁷ are both —H or bothmethyl; and X⁸ is fluoro, chloro, bromo or iodo; or (2) X⁴ isazepan-1-yl, morpholin-4-yl, pyrrolidin-1-yl, —N(CH₃)(CH(CH₃)₂) or7-azabicyclo[2.2.1]heptan-7-yl; and R⁰ is

where X⁶, X⁷ and X⁸ are as above; or (3) X⁴ is morpholin-4-yl or—N(CH₃)₂; and R⁰ is

where X⁸ is as above; or a pharmaceutically acceptable salt thereof,dispersed, in a free base equivalent amount of at least about 2.5% byweight of the composition, in a pharmaceutically acceptable carrier;wherein said active ingredient is in solid-state form and/or thecomposition further comprises, dispersed in the carrier, apharmaceutically acceptable heavier-chalcogen antioxidant (HCA) in anamount effective to inhibit oxidation of the active ingredient at athioether linkage thereof.
 2. The composition of claim 1, wherein saidactive ingredient is present in a free base equivalent amount of atleast about 5% by weight of the composition.
 3. The composition of claim1, wherein the active ingredient comprisesN-(4-(4-((2-(4-chlorophenyl)-5,5-dimethyl-1-cyclohex-1-en-1-yl)methyl)piperazin-1-yl)benzoyl)-4-(((1R)-3-(morpholin-4-yl)-1-((phenylsulfanyl)methyl)propyl)amino)-3-((trifluoromethyl)sulfonyl)benzenesulfonamide (ABT-263) or a pharmaceutically acceptablesalt thereof.
 4. The composition of claim 3, wherein the activeingredient comprises ABT-263 free base or ABT-263 bis-hydrochloride salt(ABT-263 bis-HCl).
 5. The composition of claim 3, wherein the carriercomprises excipients selected to provide sufficient bioavailability ofABT-263 to be therapeutically effective for promotion of apoptosis whenorally administered to a non-fasting human subject in need thereof in adaily dosage amount of about 200 to about 400 mg ABT-263 free baseequivalent.
 6. The composition of claim 5, wherein said sufficientbioavailability is evidenced by a bioavailability of at least about 15%in a non-fasting dog model.
 7. The composition of claim 5, wherein saidsufficient bioavailability is evidenced by one or both of (a) an ABT-263AUC₀₋₂₄ of at least about 20 μg·h/ml, and/or (b) an ABT-263 C_(max) ofat least about 2.5 μg/ml, in a single-dose non-fasting humanpharmacokinetic study at an ABT-263 free base equivalent dose of about200 to about 400 mg.
 8. The composition of claim 5, wherein saidsufficient bioavailability is evidenced by a steady-state ABT-263 C_(mm)of about 1 to about 5 μg/ml and a steady-state ABT-263 C_(max) of about3 to about 8 μg/ml in a non-fasting human pharmacokinetic study at adaily ABT-263 free base equivalent dose of about 200 to about 400 mg. 9.The composition of claim 5, wherein said sufficient bioavailability isevidenced by at least substantial bioequivalence in a humanpharmacokinetic study to a prototype formulation that consists of a 25mg/ml solution of ABT-263 bis-HCl in a mixture of 90%phosphatidylcholine+medium chain triglycerides 53/29 and 10% ethanol.10. The composition of claim 1, wherein the carrier is liquid, havingsaid active ingredient and a pharmaceutically acceptable HCA in anantioxidant-effective amount in solution or suspension therein.
 11. Thecomposition of claim 1, wherein the carrier is solid, having said activeingredient dispersed therein in solid-state form.
 12. The composition ofclaim 1, wherein said active ingredient is in amorphous or crystallineform having a D₉₀ particle size not greater than about 30 μm.
 13. Amethod for treating a disease characterized by apoptotic dysfunctionand/or overexpression of an anti-apoptotic Bcl-2 family protein,comprising orally administering to a subject having the disease thecomposition of claim 1 in a therapeutically effective daily dosageamount.
 14. The method of claim 13, wherein the disease is a neoplasticdisease.
 15. The method of claim 14, wherein the neoplastic disease isselected from the group consisting of cancer, mesothelioma, bladdercancer, pancreatic cancer, skin cancer, cancer of the head or neck,cutaneous or intraocular melanoma, ovarian cancer, breast cancer,uterine cancer, carcinoma of the fallopian tubes, carcinoma of theendometrium, carcinoma of the cervix, carcinoma of the vagina, carcinomaof the vulva, bone cancer, colon cancer, rectal cancer, cancer of theanal region, stomach cancer, gastrointestinal (gastric, colorectaland/or duodenal) cancer, chronic lymphocytic leukemia, acute lymphocyticleukemia, esophageal cancer, cancer of the small intestine, cancer ofthe endocrine system, cancer of the thyroid gland, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, testicular cancer,hepatocellular (hepatic and/or biliary duct) cancer, primary orsecondary central nervous system tumor, primary or secondary braintumor, Hodgkin's disease, chronic or acute leukemia, chronic myeloidleukemia, lymphocytic lymphoma, lymphoblastic leukemia, follicularlymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma,multiple myeloma, oral cancer, non-small-cell lung cancer, prostatecancer, small-cell lung cancer, cancer of the kidney and/or ureter,renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of thecentral nervous system, primary central nervous system lymphoma, nonHodgkin's lymphoma, spinal axis tumors, brain stem glioma, pituitaryadenoma, adrenocortical cancer, gall bladder cancer, cancer of thespleen, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastomaand combinations thereof.
 16. The method of claim 14, wherein theneoplastic disease is a lymphoid malignancy.
 17. The method of claim 16,wherein the lymphoid malignancy is non-Hodgkin's lymphoma.
 18. Themethod of claim 14, wherein the neoplastic disease is chroniclymphocytic leukemia or acute lymphocytic leukemia.
 19. The method ofclaim 13, wherein said composition comprises as the sole or first activeingredient ABT-263 or a pharmaceutically acceptable salt thereof, and isadministered in a daily dosage amount of about 50 to about 500 mgABT-263 free base equivalent.
 20. The method of claim 19, wherein saiddaily dosage amount is about 200 to about 400 mg ABT-263 free baseequivalent.